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

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.

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

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

Objective To evaluate the efficacy and safety of liraglutide ( Lira) combined with insulin aspart 30 injection in the treatment of type 2 diabetes poorly controlled with premixed insulin aspart 30 ( Asp 30 ) . Methods Seventy eight patients with type 2 diabetes mellitus without reaching glycemic targets were randomized into two groups.On the basis of the insulin aspart 30 and oral hypoglycemic drugs, the patients in the trial group were given liraglutide 0.6 mg? d-1 sc once per night for the first week, and 1.2 mg? d-1 for the following two to sixteen weeks, while the other patients in the control group received insulin detemir ( Det) sc once a night for 16 weeks, with the beginning dosage of 0.2 U? kg -1? d-1.Before and after the treatment, the body mass index ( BMI) , waist to hip ratio ( WHR) , fasting plasma glucose ( FPG) , fast-ing C -peptide ( FCP ) , 2 -hour postprandial plasma glucose (PPG 2 h), glycosylated hemoglobin A1c (HbA1c) and insulin resist-ance index(HOMA-IR) were tested.Also, the adverse drug reactions such as weight gain and hypoglycemia reaction were estimated. Results After the treatment, the level of FPG, PPG 2 h and HbA1c decreased obviously in both groups ( P<0.01 ) , while PPG 2 h in the trial group appeared a better improvement compared to the control group ( P<0.05) ,and BMI as well as HOMA-IR was improved significantly in the trial group after the treatment ( P<0.01).Both drugs were well tolerated with minor hypoglycemia, yet Lira appeared much safer than Det.Conclusion For treatment of refractory type 2 diabetes mellitus, liraglutide combined with insulin aspart 30 injection had certain clincal efficacy with minor adverse reaction.

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

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

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

OBJECTIVEThe presence of liver fibrosis in patients with beta-thalassemia major has been demonstrated to be an important negative prognostic factor. Identification of liver fibrosis in early stage would be of great value. Hyaluronic acid (HA), type III pre-collagen (PC III), collagen IV (C IV) and laminin (LN) as serum markers were widely used in the diagnosis of liver fibrosis in patients with chronic viral infections or alcoholic liver diseases. However, their values in thalassemic liver fibrosis have not been studied. This work was to determine the serum HA, PC III, C IV and LN levels in children with beta-thalassemia major and evaluate the diagnostic utility.

METHODSerum HA, PC III, C IV and LN in 49 hospitalized children with beta-thalassemia major (aged 1 - 15 years with the media age of 6.27 years) and 41 healthy children served as controls (aged 1 - 13 years with media age of 6.40 years) were detected by radioimmunoassay (RIA). Forty-five of 49 cases were performed percutaneous liver biopsies, and the histopathological fibrosis was compared with the four serum markers. The correlation and discriminate analysis were used.

RESULTSAll the serum levels of HA, PC III, C IV and LN in beta-thalassemia were significantly higher than those in controls (P < 0.01). In 36 of 45 cases, the histopathology showed liver fibrosis including stage I and stage II by biopsies with a positive rate of 80%. The serum levels of four markers increased successively with the aggravation of liver fibrosis from stage 0 to stage II, and significant correlation was observed between the level of HA or PC III and the stage of fibrosis (HA, r = 0.379, P = 0.017; PC III, r = 0.455, P = 0.04). While there was no difference between the level of C IV or LN and fibrosis (C IV, r = 0.312, P = 0.053; LN, r = 0.310, P = 0.055). Using discriminate analysis, the discriminate function of co-detection of the four markers for the diagnosis of fibrosis was 0.002 HA + 0.003 PC III + 0.002 C IV + 0.006 LN-1.859, which had a sensitivity of 93.88%, specificity of 68.29%, predictive value of positive test and negative test of 77.97% and 90.32%, respectively. Moreover, there was a significant correlation between the serum level of HA or PC III and the liver iron concentration (HA, r = 0.318, P = 0.035; PC III, r = 0.305, P = 0.044).

CONCLUSIONThe results suggest that, in beta-thalassemia major with chronic liver disease, HA and PC III showed more practical value in diagnosing liver fibrosis than the levels of C IV and LN. The combination of the four serum markers could improve the accuracy and reliability of the diagnosis. A validation study is necessary before introducing into the prediction function during the clinical practice.

Adolescent ; Biomarkers ; blood ; Child ; Child, Preschool ; Collagen Type III ; blood ; Collagen Type IV ; blood ; Female ; Humans ; Hyaluronic Acid ; blood ; Infant ; Laminin ; blood ; Liver Cirrhosis ; blood ; complications ; diagnosis ; Male ; Prognosis ; beta-Thalassemia ; blood ; complications ; pathology