A case with the diagnosis of the onset of inflammatory bowel disease (IBD) with chronic granulo-matous disease (CGD) in Children′s Hospital Affiliated to Zhengzhou University in June 2016 was chosen, and the patient′s clinical data and whole treatment process were analyzed.According to the relevant literature from Chinese and foreign databases, the clinical characteristics were analyzed and summarized, principles of diagnosis and treatment for children who had the onset of IBD with CGD.This patient was a child of 1 year and 9 months old, and the initial symptoms included repeated diarrhea and bloody stools.He was diagnosed as " ulcerative colitis" in the local hospital.After admission, the neutrophil respiratory burst test was positive.The genetic analysis result suggested that the CYBB gene was mutated, thus obtaining the diagnosis of CGD.Then, he was given prophylactic antibiotic therapy and symptomatic treatment.After the 3 months of follow-up after discharge, the patient still had intermittent diarrhea and bloody stools.CGD is a rare primary immunodeficiency disease, and current treatment methods of CGD include hematopoietic stem cell transplantation and anti-infection treatment.When IBD patients have complications other than gastrointestinal symptoms, the IBD treatment is not effective, or there are suspected parents who are married to close relatives, or with the family history of IBD, CGD should be considered.

The investigation of environmental radioactivity level is an important basic work of ecological environment protection. It can provide basic data and scientific basis for the evaluation of radioactive environment quality and the formulation of radiation safety regulations and standards. Based on many years of practical experience of environmental radioactivity level investigation, combined with relevant regulations and standards, this paper summarizes the common investigation methods in the investigation and research of environmental radioactivity level at home and abroad, summarizes a set of environmental radioactivity level investigation scheme with strong applicability, and gives detailed suggestions on the ideas, methods, technical routes and other key points of the investigation scheme. The research results of this paper can provide a reference for the preparation of environmental radioactive level investigation scheme.

Objective The radioactivity level survey was carried out in Weizhou Island area of Beihai, to comprehensively master the radiation environment status in the area, understand the natural radioactivity level and its distribution law, and provide basic data for scientific evaluation of radiation environment quality. Methods According to the relevant standards and technical specifications, the air absorbed dose rate of gamma radiation in Weizhou Island area was monitored from April 2020 to March 2021, the concentrations of radionuclides in water, soil and other environmental samples were sampled and analyzed, and the monitoring results were analyzed and discussed in combination with the regional characteristics. Results The results show that the air absorbed dose rate of gamma radiation in Weizhou Island area ranges from 0.2～122 nGy/h; the activity concentrations of ²³⁸U, ²²⁶Ra, ²³²Th, ⁴⁰K and ¹³⁷Cs in solid samples range 17.0～37.3 Bq/kg, < 0.501～29.5 Bq/kg, < 0.766～54.3 Bq/kg, 18.7～369 Bq/kg and < 0.212～1.48 Bq/kg, respectively, the concentrations of U and Th in well water and reservoir water range 0.065～0.25 μg/L, 0.046～0.079 μg/L, the activities of ²²⁶Ra, ⁴⁰K, total α and total β range 1.42～3.08 mBq/L, 0.069～0.231 Bq/L, 0.025～0.163 Bq/L, 0.082～0.572 Bq/L, respectively, the concentrations of U and Th in seawater samples range 1.81～2.25 μg/L, 0.634～0.648 μg/L, and the activities of ²²⁶Ra, ⁴⁰K, ⁹⁰Sr and ¹³⁷Cs range 9.38～19.7 mBq/L, 11.3～11.8 Bq/L、0.193～0.866 mBq/L、1.13～1.42 mBq/L. Conclusion The environmental ionizing radiation level in Weizhou island of Beihai is in the range of background fluctuation and at a relatively low level, indicating that the radiation environmental quality of Weizhou Island and its surrounding areas is good.

Objective To investigate the changes in Th17 cells and CD4+CD25+regulatory T lym-phocytes ( Treg) as well as transcription factors and cytokines relating to them in children with Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (HLH) and to analyze their role and clinical significance. Methods Thirty-two children with newly diagnosed EBV-associated HLH in the Hematology/Oncology Department of Zhengzhou Children′s Hospital from January 2012 to December 2016 were enrolled in this study. Thirty healthy children taking physical examination in the same hospital in the corresponding period were recruited as controls. Percentages of Th17 and Treg cells in peripheral blood T lymphocytes were detected by flow cytometry. Expression of RORγt and Foxp3 at mRNA level in peripheral blood mononuclear cells was detected by real-time PCR. Levels of IL-6, IL-17, IL-10 and TGF-β1 in serum samples were measured by ELISA. Results Compared with the control group, the EBV-associated HLH group showed in-creased percentage of Th17 cells [(1. 09±0. 43)％ vs (0. 39±0. 19)％, P<0. 05] and enhanced expres-sion of RORγt at mRNA level [(1. 41±0. 37) vs (0. 67±0. 13), P<0. 05], but decreased percentage of Treg cells [(3. 66±1. 13)％ vs (6. 80±1. 15)％, P<0. 05] and inhibited expression of Foxp3 at mRNA level [(15. 97±5. 11) vs (30. 23±4. 95), P<0. 05]. All of the above mentioned changes were reversed af-ter treatment (P<0. 05). Serum levels of IL-6 and IL-17 of EBV-associated HLH group were higher than those of control group, while serum levels of IL-10 and TGF-β1 were lower (P<0. 05). Conclusion Im-balanced Th17/Treg cells might play an important role in the pathogenesis of EBV-associated HLH. Cyto-kines relating to the maintenance of Th17/Treg cell balance could be used as indicators of disease develop-ment.

OBJECTIVETo investigate the changes and roles of follicular regulatory T cells (Tfr) and follicular T helper cells (Tfh) in the pathogenesis of children immune thrombocytopenia (ITP).

METHODS32 untreated ITP patients, as well as 20 healthy controls were enrolled in this study. The proportion of circulating Tfr and Tfh cells were determined by flow cytometry; real-time PCR was performed to detect the expression of transcription factors and regulatory factors of Bcl-6, c-Maf, Blimp-1 and PD-1 mRNA; ELISA was used to detect plasma concentration of IL-2, IL-6, IL-10 and IL-21.

RESULTS(1)The proportion of Tfh cells were significantly higher (P<0.05), while the Tfr cells and the ratio of tfr/Tfh cells in ITP patients were significantly lower than that in health controls (P<0.05); (2)Correlation analysis showed that the Tfr cells and the ratio of Tfr/Tfh were positively correlated with the platelet counts and negatively with the levels of PA-IgG, while the proportion of Tfh cells was positively correlated with the levels of PA-IgG and negatively with the platelet counts in peripheral blood; (3)Transcription levels of Bcl-6 and c-Maf mRNA in CD4(+) T lymphocytes cells were significantly elevated, the Blimp-1 mRNA in CD4(+) cells and PD-1 mRNA levels of Treg were lower in ITP patients in comparison with healthy controls; (4)The higher Plasma concentration of IL-21, and lower concentration of IL-2 were found in ITP patients.

CONCLUSION(1)The lower proportion of Tfr cells and higher proportion of Tfh cells, as well as the abnormal ratio of Tfr/Tfh might account for the decreased platelet counts to be further involved in the immunological pathogenesis of children ITP; (2)The changes of plasma cytokines IL-2, IL-21 in microenvironment and the over-expression of Bcl-6 mRNA, c-Maf mRNA and the lower-expression of Blimp-1 mRNA in CD4(+) T cells, and over-expression of PD-1 mRNA in Treg cells might be account for the abnormal ratios of Tfr/Tfh cells in ITP patients.

Background and purpose:High expression of excision repair cross-complementing 1 (ERCC1) is related to resistance in patients treated with platinum-containing regimens. The ERCC1 antibody 8F1 was usually used in past studies, but it was found to have no-speciifcity recently. This study aimed to investigate the predictive role of a new ERCC1 antibody 4F9 to platinum chemotherapy in non-small cell lung cancer (NSCLC) patients. Methods:Expression of ERCC1 was detected using antibody 4F9 by immunohistochemistry (IHC) in 72 NSCLC tissues. The relationship between the expression of ERCCl and the clinical pathological parameters, the efficacy of platinum chemotherapy and overall survival of patients were explored by statistical analysis. Results: The high expression of ERCCl protein was 55.5%in 72 cases. There was no signiifcant correlation between the ERCC1 expression with gender, age, pathological type, clinical stage and lymphatic metastasis (P>0.05). Patients with low expression of ERCC1 had signiifcantly higher response rates to platinum chemotherapy, longer median survival time and 2-years survival rate comparing with those with high expression of ERCC1 (62.5%vs 37.5%;22.9 vs 18.4 month;46.9%vs 37.5%), respectively (P<0.05). Conclusion:The expression analysis of ERCC1 using new ERCC1 antibody 4F9 by IHC method is helpful to assign chemotherapeutic regimen, and guide individual platinum chemotherapy for post-operation patients.

In order to confirm the existence of indoleamine 2, 3-dioxygenase (IDO) gene in swine, and to clone the novel gene followed by the molecule structure properties and expression pattern analysis, the porcine mRNA sequences homologous to human IDO were obtained from GenBank database by bioinformatics method. By using RT-PCR, the IDO gene was cloned from porcine endothelial cell line and the accuracy of the nucleic acid sequence was confirmed, and the expression pattern of the gene was detected. The three-dimensional structure model of porcine IDO was built referring to the tertiary structure of human IDO using biological sequence analysis software and database. The results showed that the porcine IDO was identified by sequencing. The nucleotide sequences were confirmed as a novel gene after submitted to Genbank. Porcine IDO was expressed in the lung, thymus, epididymis and anterior chamber with a basic level, however in peripheral blood mononuclear cells (PBMCs) the IDO gene was highly expressed. The three-dimensional structure model of porcine IDO was similar to that of human IDO. It was suggested that identification of the structure information of porcine IDO is essential to further investigate the immunologic function of the gene. Study of IDO on NK cells-mediated xenograft rejection will be a novel therapeutic target for the development of xenotransplantation.
Amino Acid Sequence ; Animals ; Base Sequence ; Cell Line ; Cloning, Molecular ; methods ; Endothelial Cells ; metabolism ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; genetics ; metabolism ; Molecular Sequence Data ; Sequence Alignment ; Swine

In order to confirm the existence of indoleamine 2, 3-dioxygenase (IDO) gene in swine, and to clone the novel gene followed by the molecule structure properties and expression pattern analysis, the porcine mRNA sequences homologous to human IDO were obtained from GenBank database by bioinformatics method. By using RT-PCR, the IDO gene was cloned from porcine endothelial cell line and the accuracy of the nucleic acid sequence was confirmed, and the expression pattern of the gene was detected. The three-dimensional structure model of porcine IDO was built referring to the tertiary structure of human IDO using biological sequence analysis software and database. The results showed that the porcine IDO was identified by sequencing. The nucleotide sequences were confirmed as a novel gene after submitted to Genbank. Porcine IDO was expressed in the lung, thymus, epididymis and anterior chamber with a basic level, however in peripheral blood mononuclear cells (PBMCs) the IDO gene was highly expressed. The three-dimensional structure model of porcine IDO was similar to that of human IDO. It was suggested that identification of the structure information of porcine IDO is essential to further investigate the immunologic function of the gene. Study of IDO on NK cells-mediated xenograft rejection will be a novel therapeutic target for the development of xenotransplantation.

Bone marrow mesenchymal stem cells (BMSCs) have been shown to be multipotent cells that possess high self-replicating capacity. The purpose of our study was to investigate the feasibility of using enteric neuron-like cells obtained by in vitro induction and differentiated from rat BMSCs for the treatment of Hirschsprung's disease (HD). Glial cell-derived neurotrophic factor (GDNF) and neurotrophin-3 (NT-3) are neurotrophic factors that play important roles in neuronal development, differentiation, survival and function. Meanwhile, GDNF mutations are a major cause of HD. In this study, BMSCs were transfected with eukaryotic expression plasmids co-expressing GDNF and NT-3, and the transfected cells displayed neuron-like changes after differentiation induced by fetal gut culture medium (FGCM). Immunofluorescence assay showed positive expression of the neuronal marker NSE and the enteric neuronal markers PGP9.5, VIP and nNOS. Reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of GDNF and NT-3 in transfected BMSCs. The present study indicates that genetically modified BMSCs co-expressing GDNF and NT-3 are able to differentiate into enteric neuronal cells and express enteric nerve markers when induced by FGCM. This study provides an experimental basis for gene therapy to treat enteric nervous system-related disorders, such as HD.

Bone marrow mesenchymal stem cells (BMSCs) have been shown to be multipotent cells that possess high self-replicating capacity.The purpose of our study was to investigate the feasibility of using enteric neuron-like cells obtained by in vitro induction and differentiated from rat BMSCs for the treatment of Hirschsprung's disease (HD).Glial cell-derived neurotrophic factor (GDNF) and neurotrophin-3 (NT-3) are neurotrophic factors that play important roles in neuronal development,differentiation,survival and function.Meanwhile,GDNF mutations are a major cause of HD.In this study,BMSCs were transfected with eukaryotic expression plasmids co-expressing GDNF and NT-3,and the transfected cells displayed neuron-like changes after differentiation induced by fetal gut culture medium (FGCM).Immunofluorescence assay showed positive expression of the neuronal marker NSE and the enteric neuronal markers PGP9.5,VIP and nNOS.Reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of GDNF and NT-3 in transfected BMSCs.The present study indicates that genetically modified BMSCs co-expressing GDNF and NT-3 are able to differentiate into enteric neuronal cells and express enteric nerve markers when induced by FGCM.This study provides an experimental basis for gene therapy to treat enteric nervous system-related disorders,such as HD.