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.

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.

A screening study for a novel tumor suppressor gene CSRP1 in Chinese origin patients with sporadic colorectal cancer / 中华普通外科杂志

Objective In previous study,we carried out refined mapping of loss of heterzygosity (LOH) on 1q31.1-32.1 and found that a minimal region of frequent deletion was located at DIS413-D1S2622,which indicated that the region could harbor a tumor suppressor gene associated with colorectal carcinoma.This study was to screen for the potential tumor suppressor gene (TSG) on D1S413-D1S2622 in Chinese origin patients with sporadic colorectal cancer.Methods 25 genes located in the D1S413-D1S2622 region were chosen and a microarray-based high throughput screening conducted in 19 sporadic colorectal cancers to identify candidate tumor suppressor genes.The relationship between expression levels of candidate genes and the clinicopathological data was analyzed.Real-time PCR was performed to validate the microarray results.Results According to the microarray-based high throughput screening,we found 4 significantly down-expressed genes,including CSRP1,LMOD1,PPP1R12B and CFHL3.There was no significant association between of CFHL3,CSRP1,LMOD1,PPP1R12B expression and the clinicopathological data.CSRP1 could be a colorectal cancer related tumor suppressor gene.CSRP1 was down-regulated in colorectal cancer.Conclusions CSRP1 might be involved in the progression of colorectal cancer.

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.

Effects of simvastatin on differentiation and apoptosis of human promyelocytic leukemia cell line NB4 / 白血病·淋巴瘤

Objective To investigate the effects of simvastatin (SV) on the proliferation,differentiation and apoptosis of human promyelocytic leukemia cell line NB4.Methods NB4 cells were incubated with SV at different concentration with or without all-trans retinoic acid (ATRA),and NB4 cells without any treatment were taken as normal control.Cells of different groups were collected at 24 h,48 h and 72 h after incubation for further detection.Morphological changes by Wright stain were performed.MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the surface CD11b expression levels,the early stage apoptosis ratio and cell necrosis ratio.Results Treated with 15 μ mol/L SV,10 μ mol/L SV and 5 μ mol/L SV respectively,with the NB4 cells growth,the cell inhibition rates gradually increased (F =7.15,P =0.000),as well as CD11b expression levels (F =3.41,P =0.014) and AnnexinVexpression levels (F =43.38,P =0.000).Furthermore the NB4 cells treated with 15 μ mol/L SV exhibited the most significant changes with cell inhibition rate of 0.96±0.02,CD11b expression level increased to (62.41±6.37) ％ and AnnexinV expression level increased to (87.38±2.94) ％ after 72 h incubation.Combination of 15 μmol/L SV with 0.5 μmol/L ATRA displayed obvious interaction for increasing CD11b expression levels (F =4.093,P =0.025),while no significant interaction for cell inhibition rates and Annexin V expression levels were observed.After 72 h incubation,the CD11b expression levels (89.46±9.13) ％ in NB4 cells treated with 15 μ mol/L SV in combination with 0.5 μ mol/L ATRA were significantly higher than those treated with ATRA (71.27±7.27) ％ and SV (62.41±6.37) ％ (t =2.71,P =0.054; t =4.37,P =0.017)' solely.Conclusion Simvastatin in vitro inhibits NB4 cell proliferation,promotes cell apoptosis,and synergistically induces cell differentiation with ATRA dose-dependently in vitro,which indicates that SV may have the effect of synergistic anti-promyelocytic potency with ATRA.

The screening and validating of PLCE1 as a new gene related to the pathogenesis of sporadic colorectal cancer / 中华普通外科杂志

Objective To screen for and validate unknown tumor suppressor genes (TSGs) in sporadic colorectal cancer (CRC) patients.Methods Through loss of heterozygosity (LOH) analysis on chromosome 10 in sporadic CRC,we have found D10S185 (10q23.31-24.33 ) exhibit a higher LOH frequency in our previous study.In present study,we screen for unknown TSGs in this region through the microarray.The expression of the new gene was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR).RT-PCR,immunohistochemistry and Western blot were done in colorectal cancer tissues with their pair-matched normal tissues in 50 cases to validate the results of microarray.Results Through the microarray-based high throughput screening,we found 4 significant down-regulated genes:PLCE1,CPEB3,NKX2-3 and SEMA4G,among them the down-regulation of PLCE1 was most significant.The results of qRTPCR were in relative agreement with the DNA microarray data.RT-PCR,immunohistochemistry and Western blot also showed that the expression of PLCE1 was at low levels in 46％ cancer tissues compared with normal tissues,more frequent in the poor differentiation tumor in patients under age 60 years (P ＜ 0.05 ).Conclusions This study demonstrated that down-regulation of PLCE1 was related to the tumorigenesis of sporadic colorectal cancer.PLCE1 might play a suppressive role in the development of colorectal cancer.

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.

Effects of HGF in Celsior solution on levels of INF-γ. IL-4 and IL-10 in a rat liver transplantation model / 中华肝胆外科杂志

Objective To explore the impact of recombinant human hepatocyte growth factor (rhHGF) in Celsior (CS) solution on the expression of INF-γ, IL-4 and IL-10 in a rat liver transplan-tation model. Methods After flushed with CS solution with addition of rhHGF (experimental group) or saline (control group), NHBD livers were stored at 4℃; for 16 h.then they were transplanted using the two-cuff technique with arterial reconstruction. The serum levels of INF-γ, IL-4 and IL-10 at lh after reperfusion were detected using ELISA. The INF-γ, IL-4 and IL-10 mRNA in the corresponding liver tissue were determined by RT-PCR. The 7-day survival rate was calculated and the histopatho-logical examination results were analyzed by hematoxylin and eosin staining. Results Compared with the control group, the experimental group showed lower INF-γ level and higher IL-4 and IL-10 levels in serum at 1 h after reperfusion (P<0. 05). The level of INF-γ mRNA in liver tissue was significant decreased at 1 h after reperfusion (P<0. 05) , and the level of IL-4 and IL-10 mRNA was significantly increased in the experimental group (P<0. 05). In experimental group, recipients got a better survival rate and histopathological examination showed a well-preserved hepatic architecture without hepatocyte necrosis, milder sinusoidal and portal congestion. Conclusion Adding exogenous rhHGF in CS solu-tion can protect NHBD livers from ischemia-reperfusion injury and prolong the survival in rats, which might be due to down-regulation of TNF-γ and up-regulation of IL-4 and IL-10.

Comparison of morphology, immunology and cytogenetics in adult acute leukemia of different ages / 白血病·淋巴瘤

Objective To investigate the characters of morphology,immunology and cytogenetics of adult acute leukemia (AL) in different ages. Methods 172 cases of newly diagnosed adult AL were divided into two groups:the non-aged group (age＜60 years) and the aged group ( age≥60 years). Morphology,immunology and cytogenetics between the two groups were compared. Results The incidence of M3 in aged AL was significantly lower than that in non-aged AL[6.0 %(3/50) vs 18.9 %(23/122),P ＜0.05]. The incidence of hypo-or extremely hypo-cellular AL in aged AL was significantly higher than that in non-aged AL[14.0 %(7/50) vs 4.1 %(5/122),P ＜0.05],but the incidence of hyper-or extremely hyper-cellular was significantly lower than that in non-aged AL[52.0 %(26/50) vs 73.8 %(90/122),P ＜0.05]. Among aged acute myeloid leukemia (AML),the incidence of lymphoid antigen positive AML (Ly+AML) was significantly higher than that in non-aged AML[63.4 %(26/41) vs 41.3 %(38/92),P ＜0.05]. The incidence of adverse karyotypes in aged AML was significantly higher than that in non-aged AML[33.3 %(11/33) vs 14.1 %(10/71),P ＜0.05].Conclusion Age is an important prognostic factor in AL. Generally,aged AL has poorer prognosis than nonaged AL.

Molecular cytogenetic abnormalities in chronic lymphocytic leukemia detected by interphase fluorescence in situ hybridization / 白血病·淋巴瘤

Objective To investigate molecular cytogenetic abnormalities in chronic lymphocytic leukemia and clinic prognostic significance. Methods Conventional cytogenetics (CC) examination was performed in 17 cases with CLL by I-FISH with five probes [DI3S25(13q14.3), ATM(11q22.3), RB1(13q14), p53(17p13.1) and CSP12(12p11.1-12q11.1)]to detect molecular cytogenetic abnormalities in CLL. Results Among 17 cases of CLL, by CC examination, only 18.75 % patient were found to have chromosomal abnormalities;whereas on I-FISH, 70.6 % patient were found to have molecular cytogenetic abnormalities including 13q-(47.1%) del(RB1) (23.5 %), del(13q13.4)(29.4 %), trisomy 12 (29.4%), del(17p13.1)(11.8 %), del (ATM)(5.6 %), the frequency of complex abnormalities were 11.8 %. No correlation of molecular cytogenetic abnormalities with sex, age, Binet stage, LDH and β_2-MG were found. Conclusion I-FISH is a more rapid, accurate and sensitive technique for detection of molecular cytogenetic abnormalities in CLL than CC, There was no statistically significant difference between molecular cytogenetic abnormalities and clinic characteristics, but its prognostic significance in CLL needs to be further investigated.

Abnormal expression of B cell activating factor in peripheral lymphocytes of some kidney transplant recipients and its potential biological significance / 中国组织工程研究

BACKGROUND:B cell activating factor belonging to tumor necrosis factor family(BAFF)is essential to B cell differentiation,maturation,survival,and antibody secretion via binding to its receptors,and may play a role in the development of T cell response.Whether or not BAFF signal participates in the kidney allograft rejection is worthy of studying.OBJECTIVE:To analyze the expression and potential bioactivity of BAFF in the peripheral lymphocytes of kidney transplant recipients.DESIGN,TIME AND SETTING:A case observation was performed at the Department of Urology,Third Affiliated Hospital of Soochow University from June 2006 to March 2007.PARTICIPANTS:Eighty-six kidney transplant recipients comprising 60 males and 26 females,aged 12-62 years old,who received first-time kidney transplantation,were included.The serum creatinine levels ranged between 65-267 μmol/L.METHODS:Peripheral blood of follow-up recipients was taken for anticoagulation using EDTA-Na2.Renal graft biopsy samples of some patients were collected.MAIN OUTCOME MEASURES:The expression rates of BAFF+,BAFF-R+,CD4+,CD8+,CD4+ CD25+ CD127-/low,CD134+,CD4+ CD 134+ and CD19+ BAFF-R+ in peripheral mononuclear cells were analyzed,and the ratio of CD4/CD8 was calculated.Biopsy tissue was subjected to pathological and immunohistochemical analyses.RESULTS:BAFF expression rate on the peripheral mononuclear cells was between 0.18%-76.97%.15%was used as the cut-off value.In the ≥15%group,the mean value of BAFF expression rate was 36.91%;BAFF+ mononuclear cells were not significantly correlated to the ratio of peripheral CD4+/CD8+ and the CD4+ CD25+ CD127-/low T lymphocytes.However,there were significant correlations between BAFF+ mononuclear cells and CD134+ lymphocytes or CD4+ CD134+ lymphocytes(P ＜0.01,P ＜0.05,respectively).While in the ＜15%group,there were no significant correlations among all indices.Pathological diagnosis confirmed that BAFF was expressed in the renal tubular epithelial cell cytoplasm and cytomembrane staining of some chronic rejection sections.CONCLUSION:Abnormal high expression of BAFF in peripheral mononuclear cells may be related to renal allograft rejection.

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.

Immunophenotype diversity of leukomonocytes cultured with fludarabine in vitro / 中国组织工程研究

BACKGROUND:Recently,some overseas in vitro researches and animal models confirmed that donor leukomonocyte cultured by fludarabine can significantly reduce acute graft-versus-host disease,but the mechanism of action is not clear.OBJECTIVE:To explore immunophenotype changes of donor leukomonocyte cultured with fludarabine in vitro.DESIGN,TIME AND SETTING:The observational cytology experiment was performed at the Department of Hematology,First People's Hospital from July 2007 to January 2008.MATERIALS:Leukomonocytes were collected from four hour healthy persons as donors of allogenic peripheral blood hematopoietic stem cell transplantation.Fludarabine was offered by Schering AG,Germany.METHODS:Peripheral blood hematopoietic stem cells collected by COBO SPECTRA were washed with RPMI1640 twice.These cells were cultured in RPMI1640 containing 0.10 volume fraction of fetal bovine serum till cell density of 2?109 L-1.1 mL of hematopoietic stem cells were washed with phosphate-buffered saline,and added CD3,CD4,CD8,CD44mAb 10 ?L,and added Mouse Ig-G1-FITC/PE as control group.These cells were attenuated with phosphate-buffered saline after incubated for 30 minutes at a low temperature,then detected by flow cytometry.Other cells were put in 24-well cultivate plate,added 8 mmol /L fludarabine as a final concentration for 24 hours(5% CO2,37 ℃).MAIN OUTCOME MEASURES:Studying immunophenotypic changes of donor leukomonocytes by flow cytometry.RESULTS:There was no significant change in the ratio of CD4+CD44+T cells before or after donor leukomonocytes cultured with fludarabine for 24 hours,and proportion of CD8+CD44+T cells declined significantly(P

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.

Loss of heterozygosity analysis to define putative region involved in tumor differentiation and metastases in sporadic colorectal cancer patients / 中华外科杂志

<p><b>OBJECTIVE</b>To detect putative suppressor loci involved in tumor progressing or metastases.</p><p><b>METHODS</b>Thirty microsatellite marker primers were employed to amply the corresponding loci of the genome DNA from 83 patients with sporadic colorectal cancer. The PCR products were electrophoresed on a 377 PRISM sequencer and the fluorescent signals were analyzed by Genotyper and Genescan software.</p><p><b>RESULTS</b>The data were obtained from 24 loci, with an average LOH frequency of 15.16%. The LOH at D2S206 and D2S364 was more frequent than 30%, and was less than 20% at the rest loci. Significant difference was observed between the percentage of LOH and tumor staging or differentiation at D2S142 (2q24.1), D2S126 (2q35), D2S2211 (2p24.2), D2S305 (2p23.3). Occarrence of deletion at the later two loci was correlative.</p><p><b>CONCLUSIONS</b>Frequent LOH was not observed at the loci around known mismatch repair genes on chr. 2. The region between D2S305 (2p23.3) and D2S2211 (2p24.2) deleted holistically, and was correlated to the stage and differentiation of tumor attended by D2S142 (2q24.1) and D2S126 (2q35) on 2q. It is suggested that unknown genes associated with tumor progressing or metastases reside in the two loci on 2q or the region on 2p.</p>

Overexpression of cyclin A in hepatocellular carcinoma and its relationship with HBx gene integration / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the frequency of cyclin A overexpression in hepatocellular carcinoma (HCC) and its relationship with clinical significance and HBx gene integration.</p><p><b>METHODS</b>PCR, RT-PCR and Western blot were used to detect the gene, mRNA and protein level of cyclin A in the tumor and nontumorous tissue. PCR and Southern blot were used to detect the integration of HBx gene in HCC.</p><p><b>RESULTS</b>Amplification of cyclin A gene was found in 1 of 35 patients; overexpression of cyclin A mRNA and protein was found in 16 of 35, 21 of 35 patients, respectively. Overexpression of cyclin A protein was correlated with patient's age, tumor size and HBx integration.</p><p><b>CONCLUSION</b>Overexpression of cyclin A occurs in the early stage of HCC carcinogenesis. It may be one of the important approaches by which HBV affects the normal cell cycle of hepatocyte.</p>

Allelic analysis on chromosome 5 in sporadic colorectal cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate and map the putative tumor suppressor loci on chromosome 5 involved in tumor progress or metastasis.</p><p><b>METHODS</b>Chromosome 5 of 83 patients with sporadic colorectal cancer was systemically screened. Fifteen microsatellite marker primers labeled with 3 different fluorescents were used to amplify the corresponding loci of the genome DNA. The PCR products were electrophoresed on a 377 PRISM sequencer and the fluorescent signals were analyzed with Genotyper and Genescan software.</p><p><b>RESULTS</b>The highest loss of heterozygosity (LOH) ratio was found at D5S416 (48.15%) on 5p and at D5S471 (38.71%) on 5q. The region (5q13.3 - 31), where D5S471 and 3 neighboring loci (D5S428, D5S2027 and D5S2115) reside, presented high frequent LOH.</p><p><b>CONCLUSION</b>The deletion of APC, MCC, CTNNA1 and IL cluster in the 5q 13.3 - 31.1 area play important role in the tumorogenesis of colorectal cancer, and the expected existence of another novel tumor suppressor gene on 5p is possible.</p>

Loss of heterozygosity of chromosome 20 in sporadic colorectal cancer / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To analyze the loss of heterozygosity (LOH) of chromosome 20 in patients with sporadic colorectal cancer to identify additional loci involved in colorectal tumorigenesis.</p><p><b>METHODS</b>Polymorphic microsatellite markers were analyzed in 83 colorectal cancer patients' tumor and normal DNA by PCR. PCR products were electrophoresed on an 377 DNA sequencer. Genescan 2.1 and Genotype 2.1 software were used in the LOH scanning and analysis. Comparisons between LOH frequency and clinicopathological data were performed by chi(2) test. P < 0.05 was considered statistically significant.</p><p><b>RESULTS</b>The average LOH frequency in the long arm, short arm and whole chromosome 20 was 21.1%, 26.7% and 22.8%, respectively. Chromosome 20 exhibited relatively high LOH frequency, particularly in the regions of 20p and 20q11.1-q13.1.</p><p><b>CONCLUSION</b>There is notable genetic instability on chromosome 20 in sporadic colorectal carcinoma patients; that is, mutation on chromosome 20 is closely associated with sporadic colorectal carcinogenesis. Also, there may be tumor suppressor genes related to sporadic colorectal carcinoma near the region 20q11.1-q13.1.</p>

Loss of heterozygosity of chromosome 22 in sporadic colorectal carcinoma / 中华消化杂志

Objective The loss of heterozygosity (LOH) on tumor suppressor genes is believed to play a key role in the carcinogenesis of colorectal cancer. When LOH occurs at a tumor suppressor gene locus where one of the alleles is already abnormal, it can result in neoplastic transformation. In this study, we analyzed the LOH on the chromosome 22 in sporadic colorectal cancer to identify additional loci involved in colorectal tumorigenesis. Methods Six polymorphic microsatellite markers were analyzed in 83 cases of colorectal cancer and normal tissue DNA by PCR. PCR products were eletrophoresed on an ABI Prism 377 DNA sequencer; Genescan 3.1 and Genotype 2.1 software were used for LOH scanning and analysis. Comparison between LOH frequency and clinicopathological data were performed by ? 2 test. Results The average LOH frequency on chromosome 22 was 27.27%. The region between markers D22S280 and D22S274 (22q12.2-q13.33) exhibited relatively high LOH frequency. The two highest LOH loci with frequencies of 35.09% and 34.04% was identified on D22S280 (22q12.2-q12.3) and D22S274 (22q13.32-q13.33). On D22S274 locus, LOH frequency of rectal cancer was 50% (9/18), which was higher than that of proximal colon cancer (12%, 2/17) ( P =0.018). The frequency of distal colon cancer was 42% (5/12), also higher than that of proximal colon cancer. But there was no statistical significance. Putting both the tumors in distal colon and rectum together into consideration, the frequency, 47% (14/30), was higher than that of proximal colon cancer ( P = 0.015 ),suggested the mechanism of carcinogenesis was different in both groups.Conclusion This study provided evidence for the involvement of putative tumor suppressor genes related to the sporadic colorectal carcinoma on chromosome 22q. The tumor suppressor gene(s) might locate on the 22q12.2-q12.3 and/or 22q13.32 -q13.33.