This study established superparamagnetic iron oxide (SPIO)-labeled nerve growth factor-beta (NGF-beta) gene-modified spinal cord-derived neural stem cells (NSCs). The E14 rat embryonic spinal cord-derived NSCs were isolated and cultured. The cells of the third passage were transfected with plasmid pcDNA3-hNGFbeta by using FuGENE HD transfection reagent. The expression of NGF-beta was measured by immunocytochemistry and Western blotting. The positive clones were selected, allowed to proliferate and then labeled with SPIO, which was mediated by FuGENE HD transfection reagent. Prussian blue staining and transmission electron microscopy (TEM) were used to identify the SPIO particles in the cells. The distinctive markers for stem cells (nestin), neuron (beta-III-tubulin), oligodendrocyte (CNPase) and astrocyte (GFAP) were employed to evaluate the differentiation ability of the labeled cells. The immunocytochemistry and western blotting showed that NGF-beta was expressed in spinal cord-derived NSCs. Prussian blue staining indicated that numerous blue-stained particles appeared in the cytoplasma of the labeled cells. TEM showed that SPIO particles were found in vacuolar structures of different sizes and the cytoplasma. The immunocytochemistry demonstrated that the labeled cells were nestin-positive. After differentiation, the cells expressed beta-III-tubulin, CNPase and GFAP. It was concluded that the SPIO-labeled NGF-beta gene-modified spinal cord-derived NSC were successfully established, which are multipotent and capable of self-renewal.
Cells, Cultured ; Dextrans/*diagnostic use ; Embryo, Mammalian ; Magnetic Resonance Imaging ; Magnetics ; Magnetite Nanoparticles/*diagnostic use ; Nerve Growth Factor/*genetics ; Nerve Growth Factor/pharmacology ; Neural Stem Cells/*cytology ; Spinal Cord/*cytology ; Transfection

Objective:To investigate the correlation between IL-10 and I1-17 expression levels in Mycoplasma pneumoniae pneumonia (MPP) and lung function.Methods:70 patients were included in this study.According to wheezing or not,they were divided into wheezing group and non-wheezing group.Another 30 healthy children were taken as a control group.After taking fasting blood 5ml,the serum IL-10 and IL-17 expression levels were detected by ELISA.The forced expiratory volume in one second (PEV1),peak expiratory flow (PEF) and forced vital capacity (FEV1 / FVC) of all subjects were detected.Results:The IL-10 expression level of the wheezing group were significantly different with that of the control group (P＜0.05) and that of the non-wheezing group (P＜0.05).The 1L-17 expression level of the wheezing group also had significant difference (P＜0.05) with that of the control group and non-wheezing group (P＜0.05).The IL-10 expression levels of wheezing and non-wheezing group all were lower than that of the control group.Whereas the IL-17 expression levels of wheezing and non-wheezing group all were higher than that in the control group.In addition,patients in wheezing group had higher PEV1,PEF,PVE1/FVC values than those in non-wheezing group,with significant difference (P＜0.05).The serum level of IL-10 expression of Mycoplasma pneumonia patients was positively correlated with PEV1,PEF and PVE1/FVC,while the serum level of IL-17 expression of Mycoplasma pneumonia patients was negatively correlated with PEV1,PEF and PVE1/FVC.Conclusion:The serum levels of IL-17 and IL-10 expression of Mycoplasma pneumonia children had close correlation with their pulmonary function.

Laparoscopic cholecystectomy is considered the gold standard for the treatment of symptomatic cholecystolithiasis and has become one of the typical representatives of minimally invasive surgery. This article briefly introduces the contraindication and indication for laparoscopic cholecystectomy, commonly used surgical procedures, and possible complications and related treatment methods, emphasizes the improvement of surgical procedure and the development of new equipment for cholecystectomy, and points out the current status, problems, and development trend of laparoscopic cholecystectomy, in order to provide a reference for better application of laparoscopic cholecystectomy in clinical practice.

Aim To investigate the impact of CYP3 A5 genetic polymorphism on modified releasing tacrolimus pharmacokinetics in Chinese stable renal transplant re-cipients. Methods Pharmacokinetics of once daily-ta-crolimus( tac-q. d. ) and twice daily-tacrolimus( tac-b. i. d. ) were determined by CLIA, CYP3A5 genotype was measured by PCR-RFLP. Each 10 patients receiv-ing tac-q. d. and tac-b. i. d. respectively were en-rolled, and each 5 patients receiving tac-q. d. were matched to poor metabolizer ( PM ) and extensive me-tabolizer ( EM ) group respectively according to CYP3A5 genotypes. Results AUC0~24 h for tac-q. d. was 1. 78 folds higher than AUC0~12 h for tac-b. i. d. , and dose-adjusted C0 was 40% lower for tac-q. d. than for tac-b. i. d. There were no significant differences for other parameters between the two groups; Cmax, AUC0~24 h and C0 were 1. 75, 1. 96 and 2. 49 folds higher for PM than for EM, and dose-adjusted Cmax, AUC0~24 h and C0 were 1. 80, 2. 34 and 2. 64 folds higher for PM than for EM. There were good correla-tions between AUC0~24 h and C0 for tac-q. d. Conclu-sion Conversion from tac-b. i. d. to tac-q. d. results in requirement of increased tacrolimus dose and detec-tion of CYP3A5 genotype, which is necessary for ensu-ring C0 in the range of therapeutic window.

Objective To explore the characteristics of morphology and immunology in acceleration phase and blastic phase of chronic myelogenous leukemia(CML).Methods Seventy-three cases of CML-BP bone marrow specimens were respectively conducted the morphology and related cell chemical dyeing observation for determining the FAB type.Flow cytometry was used to detect series immunological related antigens.Results Among 73 cases of FAB typing,there were 44 cases of CML-AML,21 cases of ALL and 8 cases.21 cases CML-ALL patients In the immunophenotyping by flow cytometry,among 21 cases of CML-ALL,there were 19 cases of B-ALL,2 cases of T-ALL,moreover 12 cases contained myeloid marker.Among 8 cases CML-HAL,the immunophenotypes were 6 cases of B+-My and 2 cases of T+ My.Among 44 cases CML-AML,15 cases contained T cell marker,and 2cases contained B cell marker,other cases had no cross-lineage expression.Among 73 cases of CML-BP,29 cases conducted the flow cytometry detection in the acceleration phase,in which 16 cases urgently changed to AML,and 13 cases to non-AML(9 cases of ALL and 4 cases of HAL).Among non-AML cases,2 cases had the simultaneous existence of myeloid primitive cells and precursor lymphocyte in the acceleration phase and other 9 cases were myeloid primitive cell or accompanied by lymphocyte marker.Conclusion Flow cytometry has a certain implication role for the direction and differentiation diagnosis of CML-BP.

This study established superparamagnetic iron oxide (SPIO)-labeled nerve growth factor-β (NGF-β) gene-modified spinal cord-derived neural stem cells (NSCs).The E14 rat embryonic spinal cord-derived NSCs were isolated and cultured.The cells of the third passage were transfected with plasmid pcDNA3-hNGFβ by using FuGENE HD transfection reagent.The expression of NGF-β was measured by immunocytochemistry and Western blotting.The positive clones were selected,allowed to proliferate and then labeled with SPIO,which was mediated by FuGENE HD transfection reagent.Prussian blue staining and transmission electron microscopy (TEM) were used to identify the SPIO particles in the cells.The distinctive markers for stem cells (nestin),neuron (β-Ⅲ-tubulin),oligodendrocyte (CNPase) and astrocyte (GFAP) were employed to evaluate the differentiation ability of the labeled cells.The immunocytochemistry and western blotting showed that NGF-β was expressed in spinal cord-derived NSCs.Prussian blue staining indicated that numerous blue-stained particles appeared in the cytoplasma of the labeled cells.TEM showed that SPIO particles were found in vacuolar structures of different sizes and the cytoplasma.The immunocytochemistry demonstrated that the labeled cells were nestin-positive.After differentiation,the cells expressed β-Ⅲ-tubulin,CNPase and GFAE It was concluded that the SPIO-labeled NGF-β gene-modified spinal cord-derived NSC were successfully established,which are multipotent and capable of self-renewal.

Severe acute pancreatitis (SAP) is closely related to intestinal mucosal barrier dysfunction, in which intestinal epithelial mechanical barrier injury is the structural basis of SAP-related intestinal mucosal dysfunction. Among the molecular mechanisms of injury factors, inflammatory mediators and cytokines, produced by SAP waterfall inflammation, are the main factors of intestinal mucosal barrier injury. This article briefly describes the effects of SAP on intestinal mechanical barrier injury and its molecular mechanism in order to provide ideas for the treatment of SAP.

【Objective】 To conduct a case-control study on precocious puberty as an example to introduce the establishment and design of the electronic Data capture and management platform using Research Electronic data Capture (REDCap) system and support the development of clinical research. 【Methods】 Based on the clinical REDCap system, the case-control research project of precocious puberty was created, the case report forms were designed, the user rights were set, and the data quality control rules were formulated. 【Results】 We established the electronic data capture and management platform for our research, which had 15 case report forms, to collect the data of the participants, including sociodemographic information, time for rest and activities, diet, exposure to environmental internal-secretion interfering-substances, physical examination and biochemical indicators. We conducted project management by setting up features such as user permissions and workgroups, and added data quality verification rules to control data quality. The data could be exported in various file formats for analysis and sharing. 【Conclusion】 The application of REDCap to establish the data capture and management platform of precocious puberty case-control study has promoted the efficient implementation of clinical research, which can be further popularized and applied to clinical researches in other fields.

Objective: To monitor the WT1 mRNA level and its dynamic changes in patients with myelodysplastic syndromes (MDS) after hypomethylating agents (HMA) , as well as to assess the significance of WT1 mRNA levels and its dynamic changes in evaluating the efficacy of HMA and distinguishing the disease status of heterogeneous patients with stable disease (SD) . Methods: Bone marrow or peripheral blood samples of 56 patients with MDS who underwent hypomethylating agents (≥4 cycles) from November 2009 to March 2018 were tested by real-time quantitative polymerase chain reaction (PCR) to detect the expression of WT1 mRNA, and to observe the correlation between the dynamic changes of WT1 mRNA expression and clinical efficacy and prognosis of patients. Results: WT1 mRNA expression levels of MDS patients decreased significantly after 3 cycles of hypomethylating agent treatment. Besides, the WT1 mRNA expression levels of patients increased significantly after diseases progression. According to the dynamic changes of WT1 mRNA expression levels during SD, 45 cases could be further divided into increased group and non-increased group. In those SD patients with increased WT1 mRNA expression level, the ratio of suffering disease progression or transformation to AML was 95.65% (22/23) , whereas the ratio turned to be 9.09% (2/22) for the non-increased group (χ²=33.852, P<0.001) . Compared with those SD patients reporting no increase in WT1 mRNA expression level, the overall survival[17 (95%CI 11-23) months vs not reached, P<0.001] and progression-free survival [13 (95%CI 8-18) months vs not reached, P<0.001] of those SD patients reporting increase in WT1 mRNA expression level were significantly shorter. Conclusion WT1 mRNA expression level is a useful indicator to assess the efficacy of hypomethylating agents in MDS patients. Especially in patients with SD, detection of the changes in WT1 mRNA expression level is able to predict disease progression and help to make clinical decision.