Clinical significances of CD4/CD8 ratio and neutrophil-to-lymphocyte ratio in patients with multiple myeloma / 白血病·淋巴瘤

Objective:To investigate the clinical significances of CD4/CD8 ratio and neutrophil-to-lymphocyte ratio (NLR) in patients with multiple myeloma (MM).Methods:The clinical data of 124 MM patients in the Third Affiliated Hospital of Soochow University from December 2002 to April 2017 were retrospectively analyzed, and 31 healthy people were chosen as the controls. Peripheral blood T lymphocyte subsets were detected by using flow cytometry, and the correlations between CD4/CD8 ration and related clinical indicators were also investigated. All MM patients were divided into the high NLR group and the low NLR group according to the media of NLR, and the correlation of them with related clinical indicators, chromosome karyotype, overall survival (OS) and progression-free survival (PFS) was also compared.Results:Compared with the healthy control group, the proportion of CD4 + T cells [(35.28±6.58)% vs. (31.85±6.76)%, t = -2.067, P = 0.043], absolute value of NK cells [0.22×10 9/L (0.13×10 9/L-0.59×10 9/L) vs. 0.17×10 9/L (0.00×10 9/L-0.42×10 9/L), Z = -2.614, P = 0.009] and CD4/CD8 ratio [0.97 (0.50-2.69) vs. 0.81 (0.30-1.28), Z = -2.253, P = 0.024] was decreased, respectively. The proportion of CD8 + cells was increased [(36.93±7.38)% vs. (40.50±6.50)%, t = 2.074, P = 0.042] in MM group. The hemoglobin level of CD4/CD8 ratio ≥0.94 group was higher than that of CD4/CD8 ratio <0.94 [(98.89±21.35) g/L vs.(80.60±23.23) g/L, t = -2.066, P = 0.047]. Compared with the healthy control group, NLR was increased in MM group [1.54 (1.10-3.23) vs. 1.95 (0.29-12.70), Z = -2.384, P = 0.017]. Compared with the low NLR group (<1.95), serum β 2-microglobulin [4.56 mg/L (1.63-12.60 mg/L) vs. 6.17 mg/L (1.58-67.50 mg/L), Z = -2.586, P = 0.010] and serum creatinine [84.5 μmol/L (43.0-376.5 μmol/L) vs. 113.0 μmol/L (46.5-754.0 μmol/L), Z = -3.866, P < 0.001] was increased in the high NLR group for MM patients. The proportion of the male patients, β 2-microglobulin > 5.5 mg/L, serum creatinine > 177 μmol/L, stage Ⅲ of international staging system (ISS) in the high NLR group was higher than that in the low NLR group (all P < 0.05), and there was no statistically significant difference in the composition of chromosome karyotype (all P > 0.05). The median OS time in the low NLR group was longer than that in the high NLR group [30 months (20-40 months) vs. 17 months (7-27 months), χ 2 = 4.519, P = 0.034], and there was no statistically significant difference in the PFS of both groups ( P > 0.05). Multivariate Cox analysis demonstrated that the age, corrected serum calcium, serum creatinine, lactic dehydrogenase were the independent influencing factors of OS in MM (all P < 0.05), while NLR wasn′t an independent influencing factor of OS in MM ( P = 0.513). Conclusions:CD4/CD8 ratio is decreased and immune dysfunction occurs in MM patients. MM patients with high NLR have a shorter OS time.

Phase analysis of gated myocardial perfusion imaging for early diagnosis of cardiotoxicity caused by anthracyclines in patients with diffuse large B-cell lymphoma / 中华核医学与分子影像杂志

Objective@#To evaluate the left ventricular systolic synchrony and investigate the early diagnostic value of left ventricular systolic dyssynchrony on cardiotoxicity caused by anthracyclines in patients with diffuse large B-cell lymphoma (DLBCL).@*Methods@#Thirty-two patients (22 males, 10 females, age: 22-73(54.4±14.2) years) from June 2016 to January 2019 with confirmed DLBCL and normal gated myocardial perfusion imaging (GMPI) before anthracyclines chemotherapy were enrolled prospectively. GMPI was performed after 6 cycles or more of chemotherapy. Changes of myocardial markers, electrocardiogram (ECG) indicators, left ventricular function indicators including left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), peak filling rate (PFR), summed motion score (SMS) and summed thickening score (STS) as well as left ventricular systolic synchrony indicators including phase bandwidth (BW), phase standard deviation (SD) and entropy before and after anthracyclines chemotherapy were analyzed. Paired t test, Wilcoxon signed rank test and χ2 test were used for data analysis.@*Results@#Compared with pre-chemotherapy, the left ventricular systolic synchrony indicators were significantly higher than those before chemotherapy (BW: (42.81±11.37)° vs (29.28±8.68)°; SD: (11.65±4.64)° vs (8.79±3.14)°; entropy: (39.84±5.51)% vs (36.19±5.94)%; t values: -9.132 to -3.173, all P<0.05). There were no significant differences in other indicators (t values: -1.161 to 1.750, z values: -1.633 to -0.096, all P>0.05). Of 32 patients, 13 patients (40.62%) had left ventricular systolic dyssynchrony, and the rate of chemotherapy-induced left ventricular systolic dyssynchrony was significantly higher than that of left ventricular dysfunction (15.62%, 5/32; χ2=4.947, P=0.025). All 5 patients with left ventricular dysfunction caused by chemotherapy had left ventricular systolic dyssynchrony. The LVEF of the chemotherapy-induced left ventricular systolic dyssynchrony group was significantly lower than that of the left ventricular systolic synchronization group ((54.54±9.25)% vs (66.79±7.65)%; t=4.087, P<0.01).@*Conclusion@#Left ventricular systolic dyssynchrony can be appeared in DLBCL patients after chemotherapy and is significantly earlier than left ventricular dysfunction, which can be an early indicator of cardiotoxicity caused by anthracycline chemotherapy.

Phase analysis of gated myocardial perfusion imaging for early diagnosis of cardiotoxicity caused by anthracyclines in patients with diffuse large B-cell lymphoma / 中华核医学与分子影像杂志

Objective To evaluate the left ventricular systolic synchrony and investigate the early diagnostic value of left ventricular systolic dyssynchrony on cardiotoxicity caused by anthracyclines in pa-tients with diffuse large B-cell lymphoma ( DLBCL) . Methods Thirty-two patients ( 22 males, 10 females, age:22-73(54.4±14.2) years) from June 2016 to January 2019 with confirmed DLBCL and normal gated myocardial perfusion imaging (GMPI) before anthracyclines chemotherapy were enrolled prospectively. GMPI was performed after 6 cycles or more of chemotherapy. Changes of myocardial markers, electrocardiogram (ECG) indicators, left ventricular function indicators including left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume ( LVEDV) , left ventricular end-systolic volume ( LVESV) , peak filling rate ( PFR) , summed motion score ( SMS) and summed thickening score ( STS) as well as left ventricular systolic synchrony indicators including phase bandwidth ( BW) , phase standard deviation ( SD) and entropy before and after anthracyclines chemotherapy were analyzed. Paired t test, Wilcoxon signed rank test and χ2 test were used for data analysis. Results Compared with pre-chemotherapy, the left ventricular systolic synchrony indicators were significantly higher than those before chemotherapy (BW: (42.81±11.37)° vs (29.28±8. 68)°;SD:(11.65±4.64)° vs (8.79±3.14)°;entropy:(39.84±5.51)％ vs (36.19±5.94)％;t values: -9.132 to-3.173, all P<0.05) . There were no significant differences in other indicators ( t values:-1.161 to 1.750, z values:-1.633 to-0.096, all P>0.05). Of 32 patients, 13 patients (40.62％) had left ventricular systolic dyssynchrony, and the rate of chemotherapy-induced left ventricular systolic dyssynchro-ny was significantly higher than that of left ventricular dysfunction (15.62％, 5/32;χ2=4.947, P=0.025). All 5 patients with left ventricular dysfunction caused by chemotherapy had left ventricular systolic dyssyn-chrony. The LVEF of the chemotherapy-induced left ventricular systolic dyssynchrony group was significantly lower than that of the left ventricular systolic synchronization group ((54.54±9.25)％ vs (66.79±7.65)％;t=4.087, P<0.01) . Conclusion Left ventricular systolic dyssynchrony can be appeared in DLBCL patients after chemotherapy and is significantly earlier than left ventricular dysfunction, which can be an early indi-cator of cardiotoxicity caused by anthracycline chemotherapy.

Changes of blood coagulation indicators in patients with lymphoplasmacytic lymphoma and their clinical significances / 白血病·淋巴瘤

Objective To observe the alteration and clinical significances of blood coagulation indicators in patients with lymphoplasmacytic lymphoma (LPL). Methods Twenty patients who were newly diagnosed LPL in the First People's Hospital of Changzhou from January 2008 to October 2017 and twenty healthy controls were studied. The patients were treated by chemotherapy, plasma exchange, supplement of coagulation factor or other supportive therapy. The parameters of prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), thrombin time (TT), D-dimer (D-D), and platelet count (Plt) were detected in LPL group and healthy controls. Results The levels of PT and APTT in LPL group were dramatically higher than those in control group [(12.9±1.2) s vs. (11.6±0.9) s, (41.7±9.8) s vs. (24.7±2.9) s], and the level of Plt in LPL group was lower than that in control group [112×109/L (3×109/L - 379×109/L) vs. 210×109/L (170×109/L - 271×109/L)], and the differences were statistically significant (all P< 0.05). There were no significant differences in FIB, TT and D-D levels between LPL group and control group (all P ＞0.05). There were no statistical differences in PT, APTT, FIB, TT, D-D and Plt levels among LPL patients with different types of immunoglobins (all P ＞ 0.05). After treatment, all the coagulation abnormalities got relieved and no patient died of hemorrhage or thrombosis. Conclusions The LPL patients have coagulation disorders and hypercoagulability, and this is independent of the type of immunoglobulin. Clinical attention should be paid to monitoring coagulation indicators to prevent the occurrence of adverse reactions.

Comparison of the efficacy and safety between flumatinib and imatinib in newly diagnosed chronic myeloid leukemia / 白血病·淋巴瘤

Objective To compare the efficacy and safety between flumatinib and imatinib in patients with newly diagnosed chronic myeloid leukemia (CML). Methods A multi-center, randomized and parallel comparison clinical trial was conducted in 24 newly diagnosed patients with Philadelphia chromosome-positive CML-chronic phase (Ph+ CML-CP) who were treated by flumatinib 400 mg/d, 600 mg/d or imatinib for 6 cycles (24 weeks). The hematology was evaluated at pre-medication and the 2nd, 4th, 6th, 8th, 10th, 12th, 16th, 20th, 24th week of post-medication. The morphology, cytogenetics and molecular biology were evaluated at pre-medication and 12th, 24th week of post-medication. Results In terms of efficacy, the main molecular remission (MMR) rate of flumatinib 600 mg/d group was higher than that of imatinib group after 24 weeks [44.44 % (4/9) vs. 14.29 % (1/7), P=0.017]. The rate of bcr-ablIS≤10 % in flumatinib 600 mg/d group was significantly higher than that in imatinib group (P=0.002). PK/PD analysis also hinted that patients treated by flumatinib 600 mg/d was more likely to get molecular reaction in the early stage compared with those treated by flumatinib 400 mg/d. In terms of safety, there was no significant difference in grade Ⅲ-Ⅳ of adverse events among flumatinib 400 mg/d group, flumatinib 600 mg/d group and imatinib group (P >0.05). The common adverse events in flumatinib group included skin toxicity, gastrointestinal reactions and diarrhea.There was no heart and cardiovascular toxicity in flumatinib group, and incidence of edema in flumatinib group was lower than that in imatinib group. Conclusions Flumatinib is a safe and effective drug for newly diagnosed patients with Ph+ CML-CP, and 600 mg/d is the appropriate clinical starting dose. Flumatinib and imatinib have similar safety in clinic.

Expression of activation-induced cytidine deaminase gene in leukemia / 白血病·淋巴瘤

Objective To investigate the characteristics of activation-induced cytidine deaminase (AID) expression level in de novo acute leukemia (AL) patients, chronic myeloid leukemia chronic phase (CML-CP), chronic myeloid leukemia blastic crisis (CML-BC) patients and leukemia cell lines. Methods The expression level of AID mRNA was measured in 89 cases of newly-diagnosed acute lymphoblastic leukemia (ALL) patients, 79 cases of de novo acute myeloid leukemia (AML) patients, 5 cases of CML-BC patients, 5 cases of CML-CP patients and leukemia cell lines NB4, THP-1, KG-1, Raji, K562 by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR), bone marrow mononuclear cells of 16 normal healthy donors were used as the control group. Results The expression levels of AID mRNA in 89 cases of ALL and 79 cases of AML were 0.006-7 463.175 and 0.005-69.107, the median expression levels were 3.785 and 1.812, the expression level of AID mRNA in the normal control group was 0.146-4.707, and the median expression level was 1.483, respectively. The AID expression levels of ALL, B-ALL, Burkitt leukemia, M4 patients and Raji cells were significantly higher than those of the normal control group (all P <0.05). Nevertheless, the AID mRNA expression levels of M3 patients and NB4, KG-1 cells were lower than those of the normal control group (all P <0.05). Furthermore, the AID mRNA expression levels of K562 cell were strikingly higher than that of the CML-CP patients (P<0.001), so were those of CML-BC, chronic myeloid leukemia myeloid blast crisis (CML-MBC), chronic myeloid leukemia lymphoblastic blast crisis (CML-LBC) patients. Conclusion AID gene shows high expression level in B-ALL, Burkitt leukemia and M4, low expression level in M3 and KG-1 cells, and obvious high expression level in CML-BC.

Significance of combined measurement of immunophenotype and international staging system in multiple myeloma / 白血病·淋巴瘤

Objective To investigate the immunophenotype characteristics,the clinical significance of CD28 and International staging system in multiple myeloma (MM).Methods BM aspirate samples from 49 newly diagnosed MM patients and 22 patients after treatment were assessed using 4 color flow cytometric analyses.These MM patients were classified according to International staging system and the consolidated risk staging system,and the survival rates and treatment efficiency of 2 staging system were compared.Results The higher stage,according to the international staging system,the lower treatment efficiency [the efficacy rates of Ⅰ,Ⅱ,Ⅲ stage were 83.3 ％ (4/6),38.5 ％ (5/13),38.5 ％ (5/13),36.5 ％ (19/52)](x2 =4.235,P =0.04),and the survival and progression-free survival time of high-risk group were significantly shortened (P ＜ 0.05).The rate of CD28 abnormal expression was no significant different between the initial and the treatment group [27 (55.1％),14 (63.4 ％)],also similar in the impact of survival and progression-free survival time (P ＞ 0.05).According to the consolidated risk staging system,the treatment efficiency,the survival and progression-free survival time of high-risk group were significantly shortened than the low ones (P =0.040).Conclusions The patients with higher ISS stage,abnormal expression of CD28,and high-risk group have poor prognosis.

Clinical significance of bcl-6, p53, c-myc aberrations in diffuse large B-cell lymphoma / 白血病·淋巴瘤

Objective To investigate aberrations of bcl-6,p53,c-myc genes in diffuse large B-cell lymphoma (DLBCL) and its clinical significance.Methods Interphase fluorescence in situ hybridization (I-FISH) was detected in 59 DLBCL patients in vivo tissue bcl-6,p53 protein,c-myc gene status.The patients were treated with CHOP or R-CHOP chemotheralpy,and the survival rates and treatment efficiency were compared.Results The p53 deletion was detected in 18 of the 59 cases (30.5 ％),bcl-6 rearrangement in 11 cases (18.6 ％),5 cases with c-myc rearrangement (8.5 ％).In the aspects of remission rate,p53 deletion positive group contained less advantage than negative ones (33.3 ％ vs 75.6 ％,x2 =9.560,P =0.002).The prognosis of bcl-6 gene rearrangement positive group different from negative group,but the difference was not statistically significant (OS,P =0.107; PFS,P =0.094),p53 deletion positive patients was in significantly worse prognosis than the negative group (OS,P =0.031; PFS,P =0.028),c-myc rearrangement positive group difference in gene rearrangement negative group,but the difference was not statistically significant (OS,P =0.163; PFS,P =0.167).In the CHOP group,prognosis of p53 deletion,c-myc rearrangement positive group were significantly worse than the negative group,the difference was statistically significant (P ＜ 0.05).In R-CHOP group,the prognostic significance of bcl-6 gene rearrangement positive group were worse (OS,P =0.003; PFS,P =0.007).Conclusion DLBCL patients with bcl-6,p53,c-myc genes aberrations are related with poor prognosis,and they can be used as prognostic factors for predicting DLBCL and guiding therapy.

Proliferative and apoptotic effects of simvastatin in combination with cytosine arabinoside on K562 cells / 白血病·淋巴瘤

Objective To investigate the effect of simvastatin (SV) in combination with cytosine arabinoside (ARA-C) on the proliferation and apoptosis of K562 cells. Methods Human K562 cells were incubated with SV and cytosine arabinoside alone or in combination and K562 cells without any treatment were taken as normal control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detections. Morphological changes by Wright stain were performed. MTT method was used to assay the growth inhibition rate and cytoflowmetry was used to detect the early stage apoptosis ratio and cell necrosis ratio. Results Compared with Ara-C group and SV group, cells in the group treated with SV combined with Ara-C showed obvious karyopyknosis,apoptosis bodies formation and significant cell growth inhibition, which were positively correlated with culture time. Combination of 15 μmol/L SV and Ara-C showed the most significant cell growth inhibition with a inhibition rate of (72±1) % at 72 h of culture, as was significantly higher than that of 15 μmol/L SV group (45±2) % and 20 μmol/L Ara-C group (44±0) % (P ＜0.01),furthermore, combination of 15 μmol/L simvastatin and Ara-C showed synergistic inhibition with Q value of 1.24 and 1.19 at 24 h and 48 h in each. The apoptosis rates at early stage (AnnexinV) detected by flow cytometry in 20 μmol/L, 15 μmol/L and 10 μmol/L SV treated K562 cells were significantly higher than that in normal K562 cells (P ＜0.01), as were positively correlated with culture time and SV dose (P ＜0.05). There were no significant difference of early apoptosis rate between the 20 μmol/L SV and 15 μmol/L SV groups (P ＞0.05), yet the very two were both higher than that of 10 μmol/L SV group (P ＜0.05). There were no statistic differences of late apoptosis rate (PI) amongdifferent treated groups (P ＞0.05). Conclusion SV inhibited K562 cell proliferation and induced cell apoptosis in vitro, and combination of SV and Ara-C exhibited obvious synergistic inhibition and apoptosis, which may increase the sensitivity of K562 cell to chemotherapy. SV at 15 μmol/L may be the best concentration for K562 cells in vitro.

Correlation between translocation of 14q32 and deletions of 13q14 in multiple myeloma / 白血病·淋巴瘤

Objective To investigate the common chromosome abnormalities in the patients with multiple myeloma and the relationships of cytogenetic abnormalities and clinical features. Methods The interphase fluorescence in situ hybridization (I-FISH) analysis method was designed to detect RB1-/13q14-and 14q32 rearrangements in 49 MM patients. The statistic value of its effect on clinical features were determined. Results FISH disclosed 14q32 translocations in 26 of the 40 (53.1%) patients. 25 out of the 49 (51.02 %) cases were found with deletion of chromosome 13q14 included del(RB1) in 9 (18.4 %) and del(13q14.3) in 18 (36.7 %). 13q14 deletion and 14q32 translocation were simultaneously observed in 18 (36.7 %) cases. Spearman correlation analysis were found associated of 14q32 rearrangement with the percentage of plasma cells in bone marrow (r=0.316, P=0.27). Conclusion The frequency of 13q14 deletion and 14q32 gene translocation in multiple myeloma are high. There is a significant correlation between the presence of 14q32 translocations and chromosome 13 abnormalities in MM patients. The percentage of 14q32 translocation in plasma cells was increased significantly. The 14q32 translocation is an independent prognostic factor.

Clinical significance of the changes of coagulation and fibrolysis parameters in malignant lymphomas / 白血病·淋巴瘤

Objective To explore the clinical significance of the coagulation and fibrolysis parameters changes for the knowledge of complicated thrombosis after chemotherapy in malignant lymphomas. Methods Morning fasting anti-coagulation blood samples were taken to detect plasma prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and thrombin time (TT) with automatic coagulation analyzer in 71 hospitalized malignant lymphomas and 20 normal controls. The plasma D-dimer levels of the two groups were detected with immunoturbidimetry. Results The levels of plasma APTT, Fib and D-dimer in 71 malignant lynphomas were (30.44±1.43) s, (3.28±0.20) g/L, (297.05±56.59) μg/L respectively, which were significantly higher than those in normal controls at the levels of (23.72±0.76) s, (2.57±0.22) g/L, (94.50±26.07) μg/L respectively (P ＜0.05). The coagulation and fibrolysis parameters were of no statistic differences between the normal controls and lymphomas of stage Ⅰ and Ⅱ (P ＞0.05). The APTT and Fib levels in lymphomas of stage Ⅲ and Ⅳ were higher than those in normal controls and lymphomas of stage Ⅰ and Ⅱ (P ＜0.05). There are 7 malignant lymphomas complicated venous thrombosis . Of the 7 cases, the FIB and D-dimer levels were higher than those of stage Ⅰ and Ⅱ, furthermore the D-dimer levels were higher than those of stage Ⅲ and Ⅳ. Conclusion The abnormalities of coagulation and fibrolysis parameters occurred in malignant lymphomas with requirement of periodic monitoring. After chemotherapy, the lymphoma patients had a high incidence of venous thrombosis, which need early prevention for prolongation of the survival.

Clinical significance of serum cholesterol levels in multiple myeloma / 白血病·淋巴瘤

Objective To investigate the relationship between serum cholesterol levels and immunoglobin types and clinical stages in the patients with multiple myeloma (MM). Methods We retrospectively analyzed the blood lipid levels in 65 patients with MM at diagnosis, including total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein Al (apo-Al) and apolipoprotein B (apo-B), and explored relationship between lipid parameters and immunoglobulin types or clinical stages in patients with MM. Thirty healthy persons were served as controls. Results Of the 65 MM patients, 53.85% were IgG type, 63.1 % were at stage Ⅲ. The levels of TC, HDL-C, LDL-C, apo Al and apo B in the patients with MM were significantly lower than that in the controls (P 0.05). Except one case of IgD type, the levels of TC, HDL-C, LDL-C, apo Al and apo B in Ig G and Ig A types of patients were significantly lower than that in the light chain type among other 64 cases (P <0.05), and TG levels in different immunoglobulin types was found no statistical differences. The levels of TC, HDL-C, LDL-C and apo A1 in the patients with stage Ⅲ were lower than that of stage I and controls (P <0.05), furthermore, the level of LDL in stage Ⅱwas lower than that in stage Ⅰ. Conclusion Hypocholesterolemia are seen in the patients with MM and serum cholesterol levels are related to MM staging.

Variation of serum ferrin, folic acid and vitamin B_(12) levels in patients with acute promyelocytic leukemia / 肿瘤

Objective:To observe the dynamic variation of serum ferritin (SF), folic acid, and vitamin B_(12) levels in patients with acute promyelocytic leukemia (APL) at different disease stages. Methods:Serum SF, folic acid and vitamin B_(12) levels were successively tested in thirty-six patients with primary APL every 1 to 3 months by using chemiluminescence analysis. Five different disease stages were selected as dynamic observation time points: first diagnosed, first complete remission (CR1), six months after CR1, relapsed stage,and CR1 for three years. Results:There were 75.0%(27/36)of patients with abnormal high levels of SF, 77.8% (28/36)of patients with abnormal low levels of folic acid, and 100%(36/36)of patients with increased vitamin B_(12) levels in first diagnosed stage. The number of patients with abnormal variations of SF, folic acid and vitamin B_(12) level was decreased in CR1 stage compared with those in first diagnosed stage (SF: P0.05). The serum SF, folic acid and vitamin B_(12) levels were in normal ranges in the patients who had 3-year CR. Conclusion:The serum SF, folic acid and vitamin B_(12) levels had dynamic variation in APL course. Increase in serum SF and vitamin B_(12) as well as decrease in folic acid are related with the active degree of APL and its tumor load.

Circulating CD4~+CD25~+ and CD8~+CD28~- T regulate cells in multiple myeloma / 中国免疫学杂志

Objective: The study was designed to evaluate the changes and significance of circulating CD4~+CD25~+ and CD8~+CD28~- regulatory T cells (Tregs) in patients with multiple myeloma (MM).Methods:CD4~+CD25~+ and CD8~+CD28~-Tregs in peripheral blood of 38 patients with MM and of 20 healthy doners were measured by flow cytometry.Serum albumin and β_2-MG in patients with MM were measured using bromocresol green method,transmission turbidimetry respectively.Results:Compared to those of the controls,the proportions of CD4~+CD25~(+/high),CD4~+CD25~(high) CD127~(low) and CD8~+CD28~-Treg cells in newly diagnosed MM patients were elevated.Furthermore,the proportions of CD4~+CD25~(high) and CD4~+CD25~(high)CD127~(low) Tregs in each clinical stage were elevated when compared to those of the controls.The number of the Tregs were increasing with clinical stages and were significantly higher in stage Ⅲ MM than in stageⅠ MM;In stageⅡand Ⅲ MM,there were also elevated proportions of CD8~+CD28~- Tregs,increasing with clinical stages.However,there were no differences when compared between stage Ⅰ MM and the controls;Both the proportions of CD4~+CD25~(+/high) and CD4~+CD25~(high)CD127~(low) Tregs in active MM were not different from stable MM,although all of them were higher than those of controls.The proportion of CD8~+CD28~- Tregs was higher in active MM than in stable MM and controls,but there were no differences when compared between active and stable MM.The proportions of both CD4~+CD25~(high) Tregs and CD4~+CD25~(high)CD127~(low)Tregs had negative correlation with the levels of serum albumin.Conclusion:MM patients have elevated levels of circulating CD4~+CD25~+ and CD8~+CD28~-Tregs,which may be an important mechanism of MM immune evasion,and may be associated with clinical stages,disease progression and prognosis of MM to some extent.

Effect of mesenchymal stem cell on the rate of Th1/Th2 with different stimulations / 基础医学与临床

Objective To investigate the effect of mesenchymal stem cell(MSC) on the balance of Th1/ Th2 with different stimulations.Methods To inoculate MSC in 24-well tissue culture plates.after 3 days, added T lymphocytes co-stimulated by PMA and Ionomycin,or the MSC were added to the two-way mixed lymphocyte culture according to different proportion.The subsets and cytokines of T lymphocytes were analyzed by flow cytomety.Results Differentiation into Th1 of T lymphocytes activated by co-stimulation can be inhibited MSC;In MLC,CD8~+T cell subsets and Th1 were evidently decreased.But Th2 cells were slightly increased.Conclusion MSC can significantly suppress CD8~(+)T cell,Th1,increase Th2.MSC has potentialities of alleviating acute graft-versus-host disease(aGVHD) and maintaining graft versus leukemia(GVL).

Investigation on specific killing acute leukemia cell reaction of the cytotoxic T lymphocyte induced by dendritic cell pulsed with frozen-thawed antigen / 生物医学工程学杂志

We investigated the effects of dendritic cell (DC) pulsed with acute leukemia cell frozen-thawed antigen on inducing the cytotoxic T lymphocyte (CTL) to get specific anti-tumor activity in vitro. DC was generated from healthy human bone marrow mononuclear cell (BMMC) in the presence of granulocyte/macrophage-colony stimulating factor(GM-CSF), interleukin-4 (IL-4) in vitro. DC pulsed with acute leukemia cell frozen-thawed antigen was co-cultured to induce T cell into specific CTL. Then we observed the effects of CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen killing acute leukemia cell specially and the influence of dendritic cell affecting the function and CD expression on CTL. The levels of CD1a, CD86, HLA-DR expression on DC pulsed with acute leukemia cell frozen-thawed antigen were obviously higher than those before culture (P<0.01). There were more CD3+CD8+ T cells in the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen, compared with those in the T cell uncultured group (P<0.01). The CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen significantly had higher activity in killing acute leukemia cell than in killing k562 cell (P<0.01), and the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen was also more effective for killing acute leukemia cell as compared with the CTL induced by DC simply, T cell co-cultured with IL-2 and T cell simply (P<0.01). The DC generated from human bone marrow mononuclear cell (BMMC) in the presence of granulocyte/macrophage-colony stimulating factor (GM-CSF), interleukin-4 (IL-4) was CD14- CD1a+CD83+DC, and it could also induce the cytotoxic T lymphocyte (CTL) to get specific anti-tumor activity in vitro. Otherwise,the increasing of CD3+CD8+ T cells in the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen implied the main role of the CD3+CD8+ T cells in the anti-tumor immunity.

ENZYME-LINKED LMMUNOSORBENT ASSAY (ELISA) FOR SOYBEAN RHIZOBIA STRAIN IDENTIFICATION / 微生物学通报

The direct ELISA was used to detect cells from pure cultures and nodules of Bradyrhizobium japonicum USDA110 and Rhizobium fredii RTt50. The optimum dilution of enzyme-linked conjugates, HRP-Ab110 and HRP-Ab50, was 1:3200 and 1:800 respectively. The optimum dilution of antibodies, Ab110 and Ab50, was 1:3200 and 1:800 separately. The optimum concentrations of antigens, USDA110 and RTt50, were both 6?10~7 cells/ml. Slow and fast-growing soybean rhizobiz can be detected and differentiated specifically by direct ELISA. Among a few strains of these two groups of soybean rhizobia, cross-reaction occurred. This was eliminated by absorption, so that specific strain could be identified by ELISA. The minimal concentration of antigen for detected was 2?10~5 cells/ml. It was found that nodules preserved by drying over silica gel or freezing were equally good, without loss in sensitivity of ELISA. ELISA was used to study the competition of USDA110 and RTt50 with indigenous rhizobia in soil pot experiment. Nodule occupacy of USDA110 ranged from 75-87.5% in different growing season of soybean and RTt50 ranged from 25-45%. The result showed that ELISA was more sensitive than agglutination.

The Standards for Moldproof Test and Their Applications for Industrial Materials and Products / 环境与健康杂志

The present article introduced the standards of moldproof test and their application in detail, expatiated on the harmful effect of the mold on industrial materials or products and the requirements of selecting the experiment parameters, discussed the limitation of the standards for moldproof test used in China.