OBJECTIVETo study the association between genetic polymorphisms of XRCC3 Thr241Met gene and susceptibility to gastric cardia and/non-cardia gastric cancer, and to investigate the combined effect between genes and surrounding environment.

METHODSA case-control study with respective control group was conducted. Genotypes were investigated by PCR-RFLP. Unconditional logistic regression models were used to estimate odd ratios (ORs) and their 95% confidence intervals(95% CI).

RESULTSThe frequency of XRCC3 CC, CT and TT genotypes were 43.2%, 46.5% and 10.3%, respectively in cardia cancer cases and 53.2%, 40.9% and 5.8% respectively in non-cardia gastric cancer cases while 59.6%, 35.1%, 5.3%, respectively in control group. Variated genotypes (CT and TT) increased the risk of cardia cancer after adjusting for potential confounders (OR = 1.76, 95% CI: 1.07-2.90). On cardia cancer risks, there seemed combined effects between variated genotype and high rate of alcohol drinking, low intake of fresh vegetables and having chronic gastritis. Combined effects between variated genotype and smoking, having chronic gastritis were observed in non-gastric cancer group.

CONCLUSIONXRCC3 variated genotype was one of the risk factors of cardia cancer while different risks of factors might exsit between cardia and non-cardia gastric cancer.

Cardia ; China ; DNA-Binding Proteins ; genetics ; Environment ; Genetic Predisposition to Disease ; Humans ; Odds Ratio ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Stomach Neoplasms ; genetics

This study was aimed to investigate the therapeutic efficacy of rabbit anti-thymocyte globulin (r-ATG) combined with cyclosporine A (CsA) and to analyse the efficacy-related factors in children with aplastic anemia (AA). Twenty five AA children treated with r-ATG [3.5 mg/(kg·d)×5 days] combined with CsA were analyzed retrospectively. The lymphocyte subgroups, CD4(+)/CD8 ratio and expression of CD55, CD59 on surface of neutrophils and erythrocytes in peripheral blood were detected by direct immunofluorescence method and flow cytometry; the responsive time, effective rate, adverse effects and infections after immunosuppressive therapy (IST) were analyzed; the distribution of T-lymphocyte subgroups in IST-effective and IST-uneffective groups was compared, and therapeutic efficacy-related factors were evaluated. The results showed that the response to treatments was found in 21 out of 25 cases, the total responsive rate was 84.0%; the response time was 3 - 6 months, average of 4 months; the effective rates in month 3, 6, 9, 12 after treatment were 56.0%, 72.0%, 80.0% and 84.0% respectively. The AA children with age ≥ 5 years old, course of disease < 6 months and absolute neutrophil value ≥ 1.5 ×10(9)/L on 30 days after IST had good curative effect; the effective rate in AA children with age ≥ 5 years old, course of disease < 6 months, high or reverse ratio of CD4(+)/CD8(+) and absolute neutrophil value ≥ 1.5×10(9)/L after IST was higher than that in AA children with age < 5 years old, course of disease ≥ 6 months, normal ratio of CD4(+)/CD8(+) and absolute neutrophil value after IST < 1.5×10(9)/L (94.4% vs 57.1%, 90.4% vs 50.0%, 94.1% vs 62.5%, 94.1% vs 62.5%) (P < 0.05). The high effective rate was observed in AA children with decrease of CD55 and CD59 expression, but there was no significant difference (P > 0.05) as compared with normal expression of CD55, CD59. It is concluded that the treatment using r-ATG (3.5 mg/kg·d × 5 d) combined with CsA is a safe and effective for children with AA. Age, course of disease and absolute neutrophil value on 30 days after IST are the main factors affecting curative affect.
Adolescent ; Anemia, Aplastic ; drug therapy ; Antilymphocyte Serum ; administration & dosage ; therapeutic use ; Child ; Child, Preschool ; Cyclosporine ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Female ; Humans ; Lymphocyte Count ; Lymphocyte Subsets ; Male ; Retrospective Studies ; Treatment Outcome

This study was aimed to investigate the effect of various cytokines on megakeryocytes expansion in vitro from human cord blood CD34(+) cells in order to establish an optimal culture system for MK expansion. Mononuclear cells were obtained by Ficoll-Hapaque density gradient separation. CD34(+) cells were positively isolated using a CD34 progenitor cell isolation kit. CD34(+) cells were placed into 24 well plates at a concentration of 2 x 10(4) per well. Each well contained 1 ml of IMDM with the present of effective MK cells growth cytokines. Clonogenic potentials of MK progenitor were assayed using a methylcellulose cultures system. The results suggested that four cytokines (IL-3 + IL-6 + TPO + FLT3L) culture system could effectively induce and expand cord blood CD41(+) MK cells. The number of CD41(+) cells expanded 154.67 +/- 32.21-fold on day 7, and 193.23 +/- 25.24-fold on day 14. In conclusion, established expansion system in vitro for MK cells provides experimental foundation for recovery of platelets after cord blood transplantation.
Antigens, CD34 ; analysis ; Blood Platelets ; cytology ; immunology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fetal Blood ; cytology ; immunology ; Humans ; Interleukin-3 ; pharmacology ; Interleukin-6 ; pharmacology ; Megakaryocytes ; cytology ; immunology ; Membrane Proteins ; pharmacology ; Platelet Membrane Glycoprotein IIb ; analysis ; Stem Cells ; cytology ; immunology ; Thrombopoietin ; pharmacology

To construct pcDNA3.1(+)/A2E eukaryotic expression vector and obtain a stable expression on HLA-I negative human K562 cell, PCR technique was employed to amplify A2E cDNA from the multi-cistron expression vector pG/A2E carrying HLA-E and HLA-A2 cDNA through internal ribozyme entry site (IRES), the cDNA was subcloned into vector pcDNA3.1(+), thus a eukaryotic expression was constructed and named pcDNA3.1(+)/A2E; then, the recombinant plasmid was transferred into the target cells, followed by screening with G418 and limiting dilution; finally, flow cytometry was adopted to detect HLA-E expression on the target cells. The results showed that HLA-E molecules were successfully expressed on K562 cells transfected with pcDNA3.1(+)/A2E (27.76%) and the expression of HLA-E molecules was not detected on K562 cells transfected with pcDNA3.1(+). It is concluded that the pcDNA 3.1(+)/A2E eukaryotic expression vector was successfully constructed and the HLA-E molecules were expressed on K562 cells. The data presented here would be expected to lay a good basis for the research of the molecular mechanism of HLA-E function and the interaction between HLA-E and the receptor on NK cells, as well as the influence of the expression of HLA-E in vitro on NK cells.
Cloning, Molecular ; DNA, Complementary ; genetics ; Eukaryotic Cells ; metabolism ; Flow Cytometry ; Gene Expression ; Genetic Vectors ; genetics ; HLA Antigens ; biosynthesis ; genetics ; HLA-A2 Antigen ; biosynthesis ; genetics ; Histocompatibility Antigens Class I ; biosynthesis ; genetics ; Humans ; K562 Cells ; Polymerase Chain Reaction ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Objective To analyze the spatial point pattern distribution characteristics of hemorrhagic fever with renal syndrome (HFRS) in Jingzhou city, Hubei province during the two seasons spring- summer and autumn-winter of 2017, to discuss its high incidence area and reason, and to provide basis for the resource allocation of public health. Methods The analytical data was collected from Infectious Disease Reporting Information System in China, and the spring-summer season was from March to August of 2017, while the autumn-winter was from the September of 2017 to the February of 2018. The Ripley's K-function and kernel density estimation were applied to analyze the spatial point pattern distribution and compare the distribution characteristics of spatial point pattern between the two seasons. Results In 2017, 133 cases of HFRS were reported in Jingzhou city, including the spring- summer and autumn-winter two pick incidences. The strongest aggregation distance was 17.77km in spring-summer season, and 14.40 km in autumn-winter season. The spatial gathering center was located in the north of Jianli County in spring-summer, and it moved to the south of Jiangling County and Shashi District in autumn-winter. Conclusions The key areas for the prevention and control of HFRS in Jingzhou City are Jiangling County, the southern part of Shashi District and the northern part of jianli county. The key groups are the residents of the urban-rural junction in the southern part of Shashi City, residents along the route of large-scale projects, and farmers engaged in agricultural planting or crayfish breeding in the gathering areas.

OBJECTIVEBased on magnetic beads based weak cation exchange chromatography (MB-WCX), matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) and ClinProTools software, the polypeptides of serum about occupational medicamentosa-like dermatitis induced by trichloroethylene (OMLDT) patients were studied, and a diagnostic model of OMLDT was built.

METHODSAccording to diagnostic criteria of OMLDT, serum of 28 OMLDT patients and 28 controls which were diagnosed by Shenzhen prevention and treatment center for occupational disease were collected. With the combination of MB-WCX and MALDI-TOF-MS, the polypeptides fingerprint of serum of 14 OMLDT patients and 14 controls were detected, what's more, the ClinProTools software and SNN algorithm was used for screening characteristic polypeptides and establishing diagnostic model of OMLDT. Then other objects were applied to validate the model to evaluate accuracy.

RESULTSA total of 159 peaks were attained by ClinProTools software, of which 33 peaks were statistical content (P < 0.05). What is more, comparing with the control group, 20 peaks in case group were decreased, and 13 peaks were increased. Two peaks of them were identified, that is 2106.29 and 3263.78, to classify and determine that two groups by receiver operating characteristic curve (ROC) analysis.2D peaks distribution map certified this finding and the area under the ROC curve was closed to 1. A model was established by SNN algorithm, whose cross validation and recognition capability were 87.5% and 98.5%, respectively. Its sensitivity and specificity were 84.8% and 82.1%, separately, which displayed good separating capacity.

CONCLUSIONIn the combination of MB-WCX, MALDI-TOF-MS and ClinProTools software, specifical different polypeptides were screened and OMLDT diagnostic model was built primarily. Also, the model and the results were positively validated, which would play a significant role in early diagnosis.

Adult ; Computational Biology ; Dermatitis, Occupational ; diagnosis ; Female ; Humans ; Male ; Software ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Trichloroethylene ; adverse effects

This study was undertaken to establish a murine model for unrelated allogeneic umbilical cord blood transplantation (UCBT). The characteristics and percentage of hematopoietic stem/progenitor cells between near-term fetal and neonatal murine peripheral blood (FNPB) and bone marrow (BM) were evaluated by flow cytometry and semisolid methylcellulose culture. BABL/c (H-2(d)) recipient mice conditioned with high dose CTX were transplanted with FNPB form C57BL/6 (H-2(b)) mice and the survival rate, hematopoietic and immunological reconstruction, graft versus host disease (GVHD) and engraftment level were observed. The results showed that the numbers of day 14 CFU-GM and CFU-GEMM in FNPB (176.40 +/- 78.39)% and (141.40 +/- 56.57)%, respectively were much higher than those in BM (75.20 +/- 26.41)% and (68.80 +/- 23.95)%, respectively. Moreover the percentage of Sca-1(+) CD34(+) cell subsets in FNPB (3.63 +/- 1.13)% was also higher than that in BM (1.41 +/- 0.8 7)%. FNPB transplantation improved survival rate and reconstituted hematopoietic and immune function in recipients. There was no evidence of GVHD. Chimeric analysis showed that the proportion of donor cells in BM of recipients was 27.94% at 21 days after transplantation. It was concluded that FNPB contains more hematopoietic stem and progenitor cells with high expansion ability and weak allogeneic immunity, which was similar to human UCB. The murine model for allogeneic UCBT (C57BL/6-->BALB/c) was established successfully.
Animals ; Animals, Newborn ; Fetal Blood ; cytology ; Flow Cytometry ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; Immunity ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Models, Animal ; Transplantation, Homologous

OBJECTIVEHemorrhagic cystitis (HC) is one of the common complications of hematopoietic stem cell transplantation (HSCT), which causes significant pain, prolongs hospitalization, and occasionally results in renal failure and death. This study aimed at investigating the incidence, risk factors, and outcome of HC in children post umbilical cord blood transplantation (UCBT) and peripheral blood stem cell transplantation (PBSCT).

METHODSFrom October 1998 to June the clinical records of 53 pediatric patients (aged 2-18 years with median age of 7.5 years) in our HCST center who underwent UCBT (n = 37) and PBSCT (n = 16) were retrospectively analyzed. Thirty out of 53 patients were diagnosed as hereditary hemolytic anemia (56.6%), and the others as haematological malignancies (43.4%): of whom 8 had acute lymphoblastic leukemia, 12 acute myeloid leukemia, 2 chronic myeloid leukemia and 1 non-hodgkin lymphoma. Conditioning regimen varied according to disease and clinical status, however based on cyclophosphamide (CTX, 120-200 mg/kg) and busulphan (BU, 12-16 mg/kg) in the cohort. Total body irradiation (TBI) or total lymphoid irradiation was added in 7 patients respectively. The patients were divided into regular treatment group (RTG) with 15 cases who received hyperhydration, alkalinizing, diuresis and Mesna during CTX infusion and prostaglandin E1 (PGE1) group (PEG) with 38 cases who received hyperhydration, alkalinizing, diuresis and Mesna plus prostaglandin E1 (0.03 microg/kg.h). The risk factors of HC were examined by univariate and multivariate analysis.

RESULTSIn all, 11 of the 53 transplanted patients developed HC (21%) with a median onset time of day +15 (rage day +2 - +25). HC was classified as early in 4 (36%) and late in 7 (64%), and scored as grade Iin 2 cases (18%), grade II in 4 (36%) and grade III in 5 (46%). There was no significant difference between RTG and PEG in the incidence of HC, however, the incidence was much higher in the group of patients who were > or = 6 years old, positive group of graft-versus-host disease (GVHD) and group of cytomegalovirus (CMV) infection than that in the group of patients who were < 6 years of age (32% vs. 8%, P < 0.05), negative group of GVHD (35% vs. 7%, P < 0.05) and CMV non-infected group (62% vs. 13%, P < 0.05), respectively. Furthermore, by multivariate analysis, > or = 6 years old (OR = 3.53, P < 0.05) and CMV infection (OR = 4.31, P < 0.05) were significant risk factors for HC. Three of 11 patients were treated with bladder irrigation. All patients recovered from HC in a median 12.8 days (range 2-53 days).

CONCLUSIONOlder age (> or = 6 years) as well as CMV infection were the risk factors of HC. Both hyperhydration and Mesna were effective in preventing HC, while addition of PGE1 could not reduce the incidence of HC. The prognosis of HC in children post HSCT was satisfactory.

Adolescent ; Age Factors ; Anemia, Hemolytic, Congenital ; metabolism ; therapy ; Body Water ; metabolism ; Child ; Child, Preschool ; Cystitis ; epidemiology ; etiology ; prevention & control ; therapy ; Cytomegalovirus Infections ; complications ; physiopathology ; Female ; Fluid Therapy ; methods ; Hematologic Neoplasms ; metabolism ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Incidence ; Male ; Mesna ; therapeutic use ; Multivariate Analysis ; Protective Agents ; therapeutic use ; Retrospective Studies ; Risk Factors ; Treatment Outcome

To evaluate the significance of FCM in minimal residual disease (MRD) detection, the immunophenotyping and leukemia-associated immunophenotypes (LAIP) of leukemia cells from 273 adult and 142 childhood patients with B lineage acute lymphoblastic leukemia (B-ALL) were detected by four to six antibody combinations of 4-color CD45/SSC gating multiparametric flow cytometry (FCM). The results showed that the B-ALL patients could be classified into 4 subtypes based on different expression CD34 and CD10: subtype I (CD34(+)/CD10(-)), subtype II (CD34(+)/CD10(+)), subtype III (CD34(-)/CD10(+)), subtype IV (CD34(-)/CD10(-)). The LAIP was observed in 100% and 92% patients of subtype I and subtype II, respectively, whereas only 79.2% in subtype III. The incidence of LAIP in total B-ALL cases was 90% by using the antibodies detected in this investigation. There was no significantce different for incidence of LAIP between adult and pediatric patients. LAIP was observed in 77.6% of patients by labeling only CD34/CD10/CD19/CD45 4-color antibody combination. It is concluded that in 90% of childhood and adult B-ALL patients LAIP can be found, which suits MRD detection by multiparameter flow cytometry.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; analysis ; B-Lymphocytes ; immunology ; Burkitt Lymphoma ; classification ; immunology ; pathology ; Cell Lineage ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; Neprilysin ; analysis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; classification ; immunology ; pathology

OBJECTIVETo compare the leukemia-associated immunophenotypes (LAIP) in patients with B lineage acute lymphoblastic leukemia (B-ALL) at diagnosis and relapse, and investigate its implications for minimal residual disease (MRD) detection.

METHODSThe immunophenotype of leukemia cells from 410 newly diagnosed and 6 relapsed patients with B-ALL were detected by four to six antibody combination, mainly CD34/CD10/CD45/CD19 of 4-color CD45/SSC gating flow cytometry (FCM).

RESULTSThe proportion of CD45 under-expressed or negative in relapsed patients was much higher than that in newly diagnosed patients, being 69.2% and 37.8% respectively. Immunophenotypic changes occurred in 9 relapsed patients (including 8 hematological relapse and 1 central nerves system relapse) when analyzed by paired samples analysis at diagnosis vs at relapse: 4 cases showed CD45 down-modulation and 2 up-modulation; 4 CD34 down-modulation and 2 CD10 up-modulation, while the expression of CD19 remained no change. MRD was observed in all 7 cases of hematological relapse 2 - 4 months before relapse, and the immunophenotype of MRD cells was the same as that in relapse.

CONCLUSIONA high frequency of immunophenotypic changes occurred at relapse and even in MRD before relapse, however the accuracy of MRD monitoring seemed not affected by the FCM strategy used in this investigation.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD19 ; immunology ; Antigens, CD34 ; immunology ; Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; methods ; Leukocyte Common Antigens ; immunology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; immunology ; pathology ; Neprilysin ; immunology ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology ; pathology ; Recurrence ; Young Adult