Impact of thoracic tumor radiotherapy on survival in non-small-cell lung cancer with malignant pleural effusion treated with targeted therapy: Propensity score matching study.

BACKGROUND: EGFR-mutant (EGFR-M) and ALK-positive (ALK-P)are common in malignant pleural effusion (MPE) with metastatic non-small-cell lung cancer (NSCLC) (MPE-NSCLC). The impact of thoracic tumor radiotherapy on survival in such patients remains unclear. We aimed to investigate whether thoracic tumor radiotherapy could improve overall survival (OS) in such patients. METHODS: According to whether or not patients accepted thoracic tumor radiotherapy, 148 patients with EGFR-M or ALK-P MPE-NSCLC treated with targeted therapy were classified into two groups: DT group without thoracic tumor radiotherapy and DRT group with thoracic tumor radiotherapy. Propensity score matching (PSM) was performed to balance clinical baseline characteristics. Overall survival was analyzed by Kaplan-Meier, compared by log-rank test, and evaluated using Cox proportional hazards model. RESULTS: Median survival time (MST) was 25 months versus 17 months in the DRT group and DT group. The OS rates at 1, 2, 3, 5 years in the DRT group and DT group were 75.0%, 52.8%, 26.8%, 11.1% and 64.5%, 28.4%, 9.2%, 1.8%, respectively (χ2 = 12.028, p = 0.001). Compared with DT group, the DRT group still had better survival after PSM (p = 0.007). Before and after PSM, factors associated with better OS through multivariable analysis were that thoracic tumor radiotherapy, radiotherapy, N0-2 , and ALK-TKIs. Grades 4-5 radiation toxicities were not observed in patients; 8 (11.6%) and 7 (10.1%) out of the DRT group suffered from Grade 3 radiation esophagitis and radiation pneumonitis, respectively. CONCLUSION: Our results for EGFR-M or ALK-P MPE-NSCLC showed that thoracic tumor radiotherapy may be crucial factor in improving OS with acceptable toxicities. Potential biases should not be neglected: Further randomized controlled trials are necessary to confirm this result.

Effect of Abdominal Circumference on the Irradiated Bowel Volume in Pelvic Radiotherapy for Rectal Cancer Patients: Implications for the Radiotherapy-Related Intestinal Toxicity.

Background: To effectively reduce the irradiated bowel volume so as to reduce intestinal toxicity from pelvic radiotherapy, treatment in the prone position with a full bladder on a belly board is widely used in pelvic radiotherapy for rectal cancer patients. However, the clinical applicable condition of this radiotherapy mode is unclear. The aim of this study was to preliminarily identify patients who were not eligible for this radiotherapy mode by analyzing the effect of abdominal circumference on the irradiated bowel volume. Methods: From May 2014 to September 2019, 179 patients with locally advanced rectal cancer were retrospectively reviewed in our center. All patients received pelvic radiotherapy. Weight, height, AC, and body mass index (BMI) were used as the research objects, and the irradiated bowel volume at different dose levels (V10, V20, V30, V40, V50) was selected as the outcome variable. Multivariate linear regression and sensitivity analyses were used to evaluate the correlation between AC and irradiated bowel volume. Generalized additive model (GAM) and piecewise linear regression were used to further analyze the possible nonlinear relationship between them. Results: Among the four body size indicators, AC showed a negative linear correlation with the irradiated bowel volume, which was the most significant and stable. In adjuvant radiotherapy patients, we further discovered the threshold effect between AC and irradiated bowel volume, as AC was greater than the inflection point (about 71 cm), irradiated bowel volume decreased rapidly with the increase in AC. t-test showed that in patients with small AC (<71 cm), the irradiated bowel volume was significantly higher than that of patients with medium-large AC (≥71 cm). Especially in patients with adjuvant radiotherapy, the mean irradiated bowel volume of patients with small AC was the highest in this study. Compared with adjuvant radiotherapy, in neoadjuvant radiotherapy, the mean difference of irradiated bowel volume between patients with medium-large AC and those with small AC was larger. Conclusion: AC is an independent factor influencing the irradiated bowel volume and has a strong negative linear correlation with it. Patients with small AC may not benefit from this common mode of radiotherapy, especially in adjuvant radiotherapy.

The effect of primary tumor radiotherapy in patients with Unresectable stage IV Rectal or Rectosigmoid Cancer: a propensity score matching analysis for survival.

BACKGROUND: To evaluate the impact of primary tumor radiotherapy on survival in patients with unresectable metastatic rectal or rectosigmoid cancer. METHODS: From September 2008 to September 2017, 350 patients with unresectable metastatic rectal or rectosigmoid cancer were retrospectively reviewed in our center. All patients received at least 4 cycles of chemotherapy and were divided into two groups according to whether they received primary tumor radiotherapy. A total of 163 patients received primary tumor radiotherapy, and the median radiation dose was 56.69 Gy (50.4-60). Survival curves were estimated with the Kaplan-Meier method to roughly compare survival between the two groups. Subsequently, the 18-month survival rate was used as the outcome variable for this study. This study mainly evaluated the impact of primary tumor radiotherapy on the survival of these patients through a series of multivariate Cox regression analyses after propensity score matching (PSM). RESULTS: The median follow-up time was 21 months. All 350 patients received a median of 7 cycles of chemotherapy (range 4-12), and 163 (46.67%) patients received primary tumor radiotherapy for local symptoms. The Kaplan-Meier survival curves showed that the primary tumor radiotherapy group had a significant overall survival (OS) advantage compared to the group without radiotherapy (20.07 vs 17.33 months; P = 0.002). In this study, the multivariate Cox regression analysis after adjusting for covariates, multivariate Cox regression analysis after PSM, inverse probability of treatment weighting (IPTW) analysis and propensity score (PS)-adjusted model analysis consistently showed that primary tumor radiotherapy could effectively reduce the risk of death for these patients at 18 months (HR: 0.62, 95% CI 0.40-0.98; HR: 0.79, 95% CI: 0.93-1.45; HR: 0.70, 95% CI 0.55-0.99 and HR: 0.74, 95% CI: 0.59-0.94). CONCLUSION: Compared with patients with stage IV rectal or rectosigmoid cancer who did not receive primary tumor radiotherapy, those who received primary tumor radiotherapy had a lower risk of death. The prescription dose (59.4 Gy/33 fractions or 60 Gy/30 fractions) of radiation for primary tumors might be considered not only to relieve symptoms improve the survival of patients with inoperable metastatic rectal or rectosigmoid cancer.

[Effect of demethylation treatment on the expression of inhibitory receptor KIR gene in NK-92MI cell line].

The aim of this study was to analyze the promoter methylation patterns of inhibitory killer cell immunoglobulin-like receptor (KIR) which gene expression and the effect of demethylation treatment were studied, and to explore the possible regulation mechanism of inhibitory kir gene expression. The promoter methylation levels of kir2DL1 and kir2DL2/kir2DL3 in NK-92MI cell line were detected by bisulfite sequencing technique. Then NK-92MI cells were treated with 5-azacytidine to induce the demethylation of CpG islands. The levels of gene expression of kir were determined by RT-PCR. The results demonstrated that the methylation frequencies of CpG dinucleotides surrounding the promoter regions of kir2DL1 and kir2DL2/kir2DL3 genes were 25% to 88% and 5% to 80% respectively. DNA-demethylating treatment with 5-azacytidine resulted in re-expression of kir2DL1 gene and increased expressions of kir2DL1, kir2DL2 and kir2DL3 genes in NK-92MI cells. In conclusion, the promoter DNA methylation participates in the regulation of kir gene expression in NK-92MI cells.

[An effective method for T-cell and B-cell simultaneous depletion in vitro from mobilized peripheral blood stem/progenitor cell graft for haploidentical transplantation].

Depletion of T and B cells from the graft is prerequisite for haploidentical transplantation to decrease the risk of GVHD and EBV-associated lymphoproliferative disease. This study was aimed to investigate the performance of T-cell and B-cell simultaneous depletion from mobilized peripheral blood stem cells (PBSCs) for the first time in China, using anti-CD3 and anti-CD19 antibodies conjugated to magnetic microbeads by the CliniMACS device. The depletion efficiency of T-cell and B-cells was analyzed by flow cytometry; the function of the stem cells after depletion was evaluated using colony assays. The results indicated that the mononuclear cell count prior to T- and B-cell depletion was 4.88 x 10(10). After depletion, the percentage of T cells was 0.02% with a log (10) depletion of 4.4. The percentage of B cells was less than 0.01% with a log (10) depletion of at least 3.3. The product contained not only CD34(+) stem cells, but also NK cells, monocytes and granulocytes. After T- and B-cell depletion the purity of CD34(+) cells was 0.98%, the number of CD34 cells was 1.84 x 10(8) and their recovery rate was 69.7%. The number of NK cells was 2.54 x 10(9) and the recovery rate of NK cells was 71.7%. In vitro colony assays showed no negative impact on function of the hematopoietic stem cells. In conclusion, the CliniMACS system can be used to efficiently deplete T and B cells from PBSCs simultaneously, without adverse effect on biological function of hematopoietic stem cells. This study provides technical platform for haploidentical hematopoietic stem cell transplantation.

In vitro study of biological characteristics of mesenchymal stem cells in patients with low-risk myelodysplastic syndrome.

The myelodysplastic syndromes (MDS) include a diverse groups of clonal and potentially malignant bone marrow disorders. Evidences exist that microenvironment cells from MDS marrow show functional abnormalities, which may be relevant to the incidence of such a disease. Mesenchymal stem cells (MSCs) are a very important component of hematopoietic microenvironment. This study was supposed to investigate the biological characteristics and functions of MSC derived from patients with MDS in low-risk. MSCs from bone marrow samples of 11 low-risk MDS patients were isolated, cultured and expanded. Morphology, immunophenotype and osteoblasts differentiation were analyzed. Their capacity of proliferation and hematopoietic supporting in vitro were measured. A real-time quantitative reverse transcriptase polymerase chain reaction method (RQ RT-PCR) was used for detecting the expression levels of relative cytokines and chemokines in MSC. MSCs from healthy donors were used as controls. The results showed that the culture-expanded cells from MDS patients displayed a typical fibroblast-like morphology. Cells were positive for SH2 (CD105), SH3 (CD73), Thy-1 (CD90), while negative for CD34 and CD45. After induction, these cells could differentiate into osteoblasts. The proliferative ability of MSCs in MDS patients were not different from those of MSC isolated from normal bone marrow (p > 0.05), however, their capacity of hematopoietic supporting in vitro were significantly weaker (p < 0.05). RQ RT-PCR detection indicated that the SDF-1 gene expression level in MSCs of low-risk MDS patients was significantly higher than that in MSC derived from healthy donors (p < 0.01). It is concluded that the abnormal function of MSC influences the regulation of hemotopoiesis in the bone marrow microenvironment of MDS patients. It is worthy to further investigate the new clue in etiological mechanism and therapeutic strategies for MDS.

[Construction and identification of Kir2ds4 RNAi lentiviral vector].

This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.

[Effects of recombinant human interleukin 11 and granulocyte colony stimulating factor in mobilization for autologous peripheral blood stem cell transplantation].

This study was aimed to evaluate the effects of recombinant human interleukin 11 (rhIL-11) and recombinant human granulocyte colony stimulating factor (rhG-CSF) in mobilization for autologous peripheral blood stem cell transplantation (APBSCT). 16 patients with non-Hodgkin's lymphoma or acute myeloblastic leukemia were given myelosuppressive chemotherapy, then were mobilized by using rhG-CSF 5 microg/(kg.d) for median 5.5 days and rhIL-11 50 microg/(kg.d) for median 4 days (experimental group) or rhG-CSF 5 microg/(kg.d) alone for median 5.5 days (control group). After mobilizing, the peripheral blood leucocyte and platelet counts, total mononuclear cells, CD34+ cells and CFU-GM counts in PBSC collection, and amount of apheresed platelet transfusion were assayed. The results showed that the peripheral blood leucocyte and platelet counts, total mononuclear cell, CD34+ cell and CFU-GM counts in PBSC collection were no significant difference between two groups (p>0.05). After APBSCT, the median time for neutrophil count>or=0.5x10(9)/L and the median time for platelet count>or=20x10(9)/L were 10.5 and 11.5 days in experimental group, while were 13 and 13 days in control group, respectively. The median amount of apheresed platelet transfusion was 3.5 unit in experimental group and 5 unit in control group. Data were significantly different between two groups (p<0.05). The adverse reactions of mobilization were mild fever, fatigue, symptoms like as common cold, poor appetite, dizziness, muscular soreness in experimental group, but only mild fever in control. These symptoms were well tolerated and overcome with drug withdrawal. It is concluded that the regimen of rhIL-11 in combination with rhG-CSF after myelosuppressive chemotherapy to mobilize PBSC is efficient and safe with rapid hematologic reconstitution and less platelet transfusions after APBSCT were used.

[Allogeneic peripheral blood hematopoietic stem cell transplantation for post-operatively treating acute non-lymphocytic leukemia patient complicated with renal cell carcinoma: one case report].

The aim of this study was to evaluate the safety and efficacy of allogeneic hematopoietic peripheral blood stem cell transplantation (allo-PBHSCT) for post-operative therapy of acute non-lymphocytic leukemia (ANLL) patient complicated with renal cell carcinoma (RCC). One ANLL patient complicated with RCC underwent an myeloablative HLA-identical relative allo-PBHSCT after RCC operation. The conditioning regimen consisted of total body irradiation, cyclophosphamide and cytarabine. Graft versus host disease (GVHD) prophylaxis regimen composed of cyclosporine A, myco-phenolate mofetil and short course of methotrexate. The results indicated that the patient achieved engraftment 16 days after transplantation with full donor-type chimerism detected by STR-PCR at 30 and 100 days after transplantation. No acute or chronic GVHD and any severe complication developed. As of March 2007, the patient remains without disease at follow-up of 44 months. In conclusion, allo-HSCT procedure is feasible and effective for post-operative therapy of ANLL patient complicated with RCC without severe toxicity.

[Quantitative change of Th cell subsets in patients with acute graft-versus-host disease and its clinical significance].

This study was purposed to investigate the change of Th cell subsets in the patients with acute graft-versus host disease (aGVHD) and to explore its role in the pathogenesis of aGVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT). 23 patients underwent allo-HSCT were selected for analysis. The aGVHD in patients was diagnosed according to clinical features, and was confirmed by skin biopsy in some patients. The peripheral blood from 23 patients was collected before and after allo-HSCT. The quantitative chenges of Th1 and Th2 cells in peripheral blood samples were detected by using flow cytometry. The results showed that out of 23 patieats the aGVHD occured in 8 patients including 1 case of grade I, 2 case of grade II, 3 cases of grade III; no aGVHD occured in 15 patients. The flow cytometry analysis revealed that the amount of Th1 cells in patients with aGVHD was much higher than that in patients without aGVHD (p < 0.01), the IFN-gamma expression of Th1 cells in patients with aGVHD of grad II - III significantly was higher than that in patients without aGVHD (p < 0.01), meanwhie the IL-4 expression of Th2 cells in patients with aGVHD of grade II - III was significantly lower than that in patients without aGVHD (p < 0.05). Dynamical detection indicated that the Th1 obviously increased before occurrence of aGVHD and before treatment of aGVHD, while the Th1 cells obviously decreased after treatment of aGVHD. The Th1 cells not changed significantly in patients without aGVHD before and after allo-HSCT. It is concluded that Th1 cells obviously increase in patients with aGVHD, this increase is related to aGVHD pathogenesis. Detecting the changes of Th cell subsets in the early stage after allo-HSCT may be contributed to early diagnosis and therapy of aGVHD.

[Late-onset noninfectious pulmonary complications after allogeneic peripheral blood stem cell transplantation].

The aim of study was to explore the incidence, risk factors, outcome and efficacious treatment of late-onset noninfectious pulmonary complications (LNIPC) after allogeneic peripheral blood stem cell transplantation (allo-PBSCT). Seventy patients received allo-PBSCT were analyzed retrospectively. The results showed that 9 out of 63 patients surviving more than 3 months occurred late-onset noninfectious pulmonary complications (14.3%). Five out of the 9 patients developed secondary pulmonary infections. In 4 patients, LNIPC caused death directly. Advanced stage of disease at transplantation and extensive chronic graft-versus-host disease (GVHD) happened in association with LNIPC. However, other transplantation-related factors including age at transplantation, gender of patient, conditioning regimen, HLA matching and GVHD prophylaxis were not significantly correlated with the incidence of LNIPC. It is concluded that performing pulmonary function test (PFT) and thoracic computer tomography should be taken routinely after transplantation. Most patients who get correct and early diagnosis for LNIPC will show a positive response to prednisone with or without CsA.

[Application of chimerism analysis to allogeneic hematopoietic stem cell transplantation by STR-PCR].

The aim of this study was to analyze chimerism, evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) by multiple short tandem repeat (STR) amplification using fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis. Peripheral blood and bone marrow in 27 patients who received myeloablative allogenetic cell transplantation were collected before and after transplantation in different times. 10 and 7 different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus/Cofiler plus PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype soft ware were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient. The results showed that donor chimerism was similar by the two methods. The median number of informative alleles was 6.3 (4 - 9) by Profiler Plus and 4.9 (2 - 6) by Cofiler Plus. The donor alleles appeared in 26 patients on day 28 post transplantation. One patient was not observed to appear donor alleles. 14 patients with 100% donor chimerism (DC) had stable engraftment and they still survive in free leukemia. 9 patients had unstable mixed chimerism (DC: 0% - 90.2%), and 5 of them relapsed after allo-HSCT, 6 patients died. Decrease of donor chimerism appeared prior to graft rejection and disease relapse. The incidence of GVHD was higher in group of full donor chimerism. It is concluded that dynamic monitoring donor chimerism by STR-PCR in combination with all auto-capillary electrophoresis is a valuable tool for predicting graft rejection, disease relapse and occurrence of GVHD, and provides a basis for early clinical intervention in the patients received allo-HSCT.

[Clinical study on pulmonary complications after allogeneic peripheral blood stem cell transplantation].

OBJECTIVE: To explore the incidence, pathogenesis, risk factors and effective treatment of pulmonary complications after allogeneic peripheral blood stem cell transplantation (allo-PBSCT). METHODS: Pulmonary complications in 70 patients received allo-PBSCT were analyzed. RESULTS: Thirty one episodes were observed in 26 patients. Among them episodes were infectious complications, including bacteria pneumonia, pulmonary fungus disease, CMV interstitial pneumonia and tuberculosis, some cases were caused by two pathogens, and 11 episodes were noninfectious complications, including late-onset noninfectious pulmonary complications (LONIPCs) (n=9), pulmonary edema (n=1) and interstitial pneumonia (n=1). The overall mortality was 12.9%. Graft-versus-host disease (GVHD) prophylaxis without MTX, severe acute GVHD and extensive chronic GVHD were high risk factors for pulmonary complications and advanced disease at transplantation, extensive chronic GVHD were significantly associated with the incidence of LONIPCs. CONCLUSION: Pulmonary disease is the main complication occurred in patients undergoing allo-PBSCT. It is of greatly importance to treat pathogens specifically and diagnose LONIPCs in its early stage.

[Id4 gene methylation for detection of minimal residual disease in acute leukemia].

OBJECTIVE: To evaluate the possibility of Id4 gene promoter methylation as a biomarker for minimal residual disease (MRD) detection in acute leukemia. METHODS: Methylation specific-PCR technique was used to detect Id4 gene methylation in samples with different percentages of leukemia cells from leukemia cell line and bone marrows from leukemia patients in complete remission (CR). RESULTS: Id4 gene methylation could be detected in samples containing 1% or lower leukemia cells. Frequency of Id4 gene methylation in acute lymphoblastic leukemia (ALL) patients in CR was 64.3% being higher than that in acute myeloid leukemia (AML) patients in CR. In 14 ALL patients with Id4 gene methylation, 8 relapsed in 12 months, while only one relapsed in 9 patients without Id4 gene methylation. In 8 AML patients with Id4 gene methylation, 5 relapsed in 12 months, while two relapsed in 20 AML patients with Id4 gene methylation. CONCLUSION: Id4 gene promoter methylation is a candidate of biomarker for MRD detection in acute leukemias.

[Changes of IL-2, IL-8, IL-10 and TNF-alpha levels in sera of patients with acute graft-versus-host disease].

The purpose of this study was to explore the roles of cytokines IL-2, TNF-alpha, IL-10 and IL-8 in the pathogenesis of acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation. The incidence of aGVHD was observed in 33 patients undergoing allogeneic hematopoietic stem cell transplantation. The aGVHD was clinically diagnosed. Sera from the 33 patients were taken before and after allogeneic hematopoietic stem cell transplantation. The IL-2, TNF-alpha, IL-8, IL-10 levels in serum of 33 patients were measured serially by using radioimmuno-assay (RIA). aGVHD occurred in 13 patients including 8 patients with aGVHD I and 5 patients with aGVHD II-IV. The results showed that the circulating levels of IL-2 and TNF-alpha were markedly elevated during aGVHD and strongly correlated with the severity of aGVHD as compared with patients without aGVHD. However, the level of the IL-10 in patients with aGVHD was significantly lower than that in patients without aGVHD. The change of IL-8 level was not significant statistically. It is concluded that IL-2 and TNF-alpha may play important roles in the pathogenesis of aGVHD, and measurement of serum IL-2 and TNF-alpha levels after allo-HSCT can provide predictive indicator for acute GVHD.

[Unrelated donor peripheral blood stem cell transplantation for hematologic malignancies].

This study was aimed to explore feasibility and efficacy of unrelated donor peripheral blood stem cell transplantation (UD-PBSCT) in treatment of hematologic malignancies. Ten patients with hematologic malignancies underwent high resolution DNA based typing HLA-matched or 1 locus mismatched UD-PBSCT. Busulfan, cyclophosphamide, Ara-C, MeCCNU and antithymocyte globulin (ATG) were used for preconditioning regimen in all cases. All patients received mycophenolate mofetile, cyclosporin A and short-term methotrexate with CD25 antibody as the graft-versus-host disease (GVHD) prophylaxis. The results showed that rapid engraftment was observed in all cases who presented full donor chimerism at 28 days post transplantation by STR-PCR. The median time of neutrophil recovery > 0.5 x 10(9)/L, platelet recovery > 20 x 10(9)/L was 13, 17.5 days respectively after transplantation. The incidence of acute GVHD was 3 cases (one case with grade I was recovered from GVHD by himself, one case with grade III was cured, one case with grade VI was died). It is concluded that above-mentioned preconditioning regimen and GVHD prophylaxis are effective approaches for unrelated donor peripheral blood stem cell transplantation in hematopoietic malignancies.

[Effects of mobilization with medium dose of rhG-CSF on the immunocyte component of peripheral blood in donors].

To investigate the changes of donor's peripheral blood immunocytes after mobilization with medium-dose recombinant human granulocyte-colony stimulating factor (rhG-CSF), the amounts of immunocytes in peripheral blood cells and the immunocyte components of donor peripheral blood mononuclear cells (PBMNC) in 12 healthy donors were detected by flow cytometry before and after mobilization with rhG-CSF 10 microg/(kg.day). The results showed that the median amounts of peripheral blood leukocytes before mobilization was 6.25 (4.7-7.8) x 10(9)/L, for lymphocytes it was 2.07 (1.63-3.10) x 10(9)/L, and for monocytes it was 0.163 (0.078-0.414) x 10(9)/L. In the fifth day after mobilization, the median amounts of peripheral blood leukocytes was 37.47 (24-72.57) x 10(9)/L, and for lymphocytes it was 3.22 (1.46-5.31) x 10(9)/L, and for monocytes, it was 1.2 (0.706-3.627) x 10(9)/L. The average amount of leukocytes after mobilization was 6.26 +/- 2.14 multiple of that before mobilization (P < 0.01), and the median amounts of lymphocytes after mobilization was 1.45 +/- 0.76 multiple of that before mobilization (P < 0.05), and the amount of monocytes after mobilization was 7.48 +/- 4.41 multiple of that before mobilization (P < 0.01). The median percentage of CD3(+) T lymphocytes before mobilization was 46.96% [(32.36-57.45)%], but 40.94% [(25.31-48.9)%] after mobilization. The ratio of CD4(+)/CD8(+) before mobilization was 1.27 +/- 0.46, while 1.36 +/- 0.51 after mobilization. The median percentage of CD4(+)CD8(+) T lymphocytes was 0.41% [(0.16-1.51)%], and 0.49% [(0.09-2.0)%] after mobilization. The median percentage of CD16(+)CD56(+) NK cells was 13.98% [(4.08-25.08)%] versus 16.65% [(12.06-33.05)%] after mobilization. The median percentage of CD3(+)CD16(+)CD56(+) NK-T cells was 2.75% [(0.37-6.38)%], but 3.13% [(0.46-5.95)%] after mobilization. The median percentage of CD20(+) B cells was 9.28% [(5.97-16.33)%], while 9.94% [(7.36-20.41)%] after mobilization. The median percentage of CD14(+) monocytes was 12.48% [(3.54-19.35)%] versus 29.52% [(16.51-36.76)%] after mobilization. The percentage of CD3(+) T lymphocytes, CD4(+)CD8(+) T lymphocytes, NK cells, NK-T cells and B lymphocytes in PBMNC did not change markedly before and after mobilization with middle-dose rhG-CSF. The ratio of CD4(+)/CD8(+) did not change significantly (P > 0.10). The percentages of CD14(+) monocytes in PBMNC after mobilization increased up to 2.87 +/- 1.51 higher than that before mobilization (P < 0.05). It is concluded that the changes of the CD14(+) monocytes after mobilization with rhG-CSF may be involved in graft rejection and graft versus host disease after allo-PBSCT.

[Detection of engraftment evidence after allogeneic hematopoietic stem cell transplantation by STR-PCR].

The purpose of this study was to observe the chimera status of 15 patients received allogeneic hematopoietic stem cell transplantation by using STR-PCR method. DNA from peripheral blood or bone marrow of donors and recipients in different time were extracted, 5 STR loci with high polymorphism were amplified by PCR. The PCR products were analyzed by PAGE and silver staining. The results showed that 15 patients had different level of engraftment. 10 patients displayed complete chimerism, five patients showed mixed chimerism. 10 patients were keeping continuance of remission, 4 patients died and one patient relapsed but still alive. The decrease of donor DNA amounts in mixed chimerism foreshowed the early graft rejection or relapse. Incidence of I-II degree aGVHD high significantly correlated with mixed chimerism. It is concluded that STR-PCR is the sensitive and accurate method for analyzing the chimera status after allogeneic hematopoietic stem cell transplantation. Mixed chimerism is a prophetic role for leukemia relapse and chimera status guides the treatment.

[Peripheral stem cell mobilization with medium dose of G-CSF in normal donors].

The study was aimed to investigate the mobilization effect of medium dose of granulocyte colony stimulating factor (G-CSF) in allogeneic peripheral stem cell transplantation and changes of T lymphocyte subgroup in PBMNC before and after mobilization. G-CSF was administered at 600 microg/d (i.e. 300 microg i.v. twice a day) for successive 5 days to 31 matched sibling or unrelated donors for the mobilization. Stem cells were harvested on the fourth day. FACS was used to analyze the NC, MNC and T lymphocyte subgroups. The results showed that the number of NC, MNC, CD34(+) cells and CFU-GM in dose of 600microg/d significantly increased (P < 0.05), compared with 300 microg/d; the time for hematological reconstruction was significantly shortened (P < 0.05); the ratio of adverse effects was not obviously increased (P > 0.05) and the median percentage of CD3(+) lymphocytes before mobilization was 46.96% [(32.36-57.45)%], but 40.94% [(25.31-48.9)%] after mobilization, while the ratio of CD4(+)/CD8(+) did not significantly changed. It is concluded that the administration of G-CSF 600 microg/d in allo-PBSCT has a good effect in the mobilization of PBSC with minor side effects, which can markedly promote hematopoietic reconstitution after transplantation. The relative amount of CD3(+) lymphocytes significantly decreased and the ratio of CD4(+)/CD8(+) remained unchanged, which may lead to alleviation of a GVHD after PBSCT.

[Unrelated HLA-matched donor marrow transplantation for one case of myelodysplastic syndrome].

To explore feasibility and efficacy of unrelated HLA matched donor marrow transplantation in treatment of myelodysplastic syndrome, one case (male, 31 years old) of myelodysplastic syndrome-refractory cytopenia with multilineage dysplasia (MDS-RCMD) has been received unrelated HLA-matched donor transplantation. Busulfan, cyclophosphamide, Ara-C, MeCCNU and antithymocyte globulin (ATG) were used for preparative regimen. Mycophenolate mofetile, cyclosporin A and short-term methotrexate were used for graft-versus-host disease prophylaxis. The results showed that neutrophil of > 0.5 x 10(9)/L, platelet of 58 x 10(9)/L and hemoglobin of 114 g/L were observed at 10, 20 days and 3 months respectively post transplantation. Disease-free survival without GVHD was 9 months. In conclusion, unrelated HLA matched donor marrow transplantation is an effective approach for treatment of patients with MDS.