Objective To establish oxidative stress model of hydrogen peroxide treatment by using human umbilical vein endothelial cells (ECS) as cell model to study the protective mechanism of anti oxidative stress and determine the signal transduction pathway of remifentanil.Methods Primary cultured human umbilical vein endothelial cells which were incubated with 0.1 M hydrogen peroxide to establish injury model to study remifentanil protection and related pathways.The experiment was divided into nine groups: control group (group C), Hydrogen peroxide group (group H1), Hydrogen peroxide+SP600125 group (group H2), Hydrogen peroxide+SB203580 group (group H3), Hydrogen peroxide+PD98059 group (group H4), hydrogen peroxide+remifentanil group (group HR1), hydrogen peroxide+remifentanil+SP600125 group (group HR2), hydrogen peroxide+remifentanil+SB203580 group (group HR3), hydrogen peroxide+remifentanil+PD98059 group (group HR4).Groups H1, H2, H3 and H4 only performed MAPK pathway blockade experiments.Groups HR1, HR2, HR3 and HR4 individually added remifentanil 10 ng/ml to protect 1 h.SOD activity, MDA level, Caspase-3 activity were detected and anti oxidative stress of remifentanil observed to confirm preliminary transduction pathway;Using RT-PCR expression levels of c-Jun before was observed before and after treated with remifentail 10 ng/ml.The aim was to determine the transduction pathway of the signaling molecules.Results Compared with group C, SOD activity were decreased significantly, MDA performance level were increased significantly in groups H1, H2, H3 and H4 (P<0.05).Compared with group H1, SOD activity was increased significantly, MDA performance level was decreased significantly in group HR1 (P<0.05).SOD activity difference and MDA performance level of groups HR2 and H2 had no statistical significance.Compared with group H3, SOD activity was increased significantly, MDA performance level was decreased siginificantly in group HR3 (P<0.05).Compared with group H4, SOD activity was increased significantly, MDA performance level was decreased significantly in group HR4 (P<0.05).Caspase-3 activity of groups H1, H2, H3 and H4 were higher significantly than that of group C (P<0.05).The level of C-Jun mRNA in group H1 was significantly higher than that of group C;But it was higher in group HR1 than that of group C, it was significantly lower than that of group H1 (P<0.05).Conclusion By activating the JNK pathway and its downstream signaling molecule c-Jun, remifentanil 10 ng/ml has the effect of increasing SOD activity, reducing the level of MDA expression and playing a role in anti oxidative stress.

Background and purpose:Three-dimensional power Doppler angiography (3D-PDA) is a new technique to investigate the vessels in the organs, but the research in thyroid is limited. The purpose of this research was to investigate three-dimensional power Doppler angiography (3D-PDA) in differentiating malignant from benign thyroid nod-ules.Methods:This study prospectively evaluated 103 lesions in 94 patients who were scheduled for surgery. The patients underwent preoperative 3D-PDA scanning. Analysis of the 3D-PDA characteristics includes blood flow pattern, the num-ber of blood vessels, the shape of vessels, the spatial distribution of the vessels, the existence of rich local blood flow within nodules or in the parenchyma surrounding the nodules. This study also analyzed the difference between the benign lesions and the malignant lesions.Results:There were 50 benign lesions and 53 malignant lesions. The sensitivity and specificity of irregular vessels, the asymmetry spatial distribution, rich local blood flow within nodules or in the parenchyma surround-ing the nodules were 64.2%, 96.0%; 56.0%, 88.0%; 54.7%, 96.0%; 60.4% and 94.0%, respectively. The sensitivity, speci-ficity, positive predictive value, negative predictive value and accuracy of 3D-PDA were 83.0%, 94.0%, 93.6%, 83.9% and 90.3%, respectively.Conclusion:3D-PDA provides a useful tool to investigate vascularization of thyroid leisions.This technique is feasible for clinical application and plays an important role in diagnosis of thyroid nodules.

OBJECTIVETo compare the difference in the long-term efficacy between all-trans retinoic acid (ATRA) combined Compound Huangdai Tablet and ATRA combined methotrexate (MTX) and 6-mercaptopurine (6MP) as the sequential maintenance treatment of acute promyelocytic leukemia (APL) patients.

METHODSTotally 83 APL patients in the molecular remission (PML/RARalpha negative) were randomly assigned to two groups, the treatment group (45 cases) and the control group (38 cases) after they were induced to the complete remission (CR) by ATRA combined chemotherapy, and treated by sequential chemotherapy as the consolidated treatment for 3 therapeutic courses. Those in the treatment group were sequentially treated with ATRA and Compound Huangdai Tablet as maintenance therapy, while those in the control group were treated with ATRA and MTX + 6MP as maintenance therapy. After a long-term follow-up (2003 -2011), the long-term therapeutic efficacy and adverse reactions were compared between the two therapeutic regimens.

RESULTSThe 5-year relapse-free survival (RFS) rate was 84.4% +/- 5.4% in the treatment group and 63.2% +/- 7.8% in the control group, showing statistical difference between the two groups (P < 0.05). The 5-year overall survival rate (OSR) was 86.7% +/- 5. 1% in the treatment group and 78.7% +/- 6.7% in the control group, showing no statistical difference between the two groups (P > 0.05). There was no statistical difference in the adverse reaction between the two groups (P > 0.05).

CONCLUSIONThe application of ATRA and Compound Huangdai Tablet as maintenance therapy could elevate the long-term RFS rate of APL patients.

Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; therapeutic use ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; Male ; Mercaptopurine ; administration & dosage ; therapeutic use ; Methotrexate ; administration & dosage ; therapeutic use ; Middle Aged ; Phytotherapy ; Treatment Outcome ; Tretinoin ; administration & dosage ; therapeutic use ; Young Adult

Acute myeloid leukemia (AML) is a genetic heterogeneous disease in which primordial and juvenile myeloid cells proliferate or accumulate abnormally in bone marrow, peripheral blood and other tissues, resulting in damage to normal hematopoietic function. Studies have shown that about 30% of AML patients have FMS-like tyrosine kinase 3 (FLT3), FLT3 abnormal regulation is closely related to the occurrence and development of AML. At present, FLT3 has become an important target for developing small molecular targeted drugs. Currently, a variety of FLT3 inhibitors and FLT3 degraders have been developed targeting FLT3, and some compounds have exhibited good anti-AML activity. This article summarizes and sorts out the current mainstream drugs for AML therapeutic targeting FLT3, in order to provide a reference for the development and design of AML drugs.

Objective:To study the correlation between systemic immune inflammation index (SII) and prognosis of patients with hilar cholangiocarcinoma after surgical treatment.Methods:The clinical data of 181 patients with hilar cholangiocarcinoma treated by surgery at the First Affiliated Hospital of Zhengzhou University from January 2012 to December 2016 were retrospectively analyzed. There were 119 men and 62 women, with an average age of 62.4 years. SII was calculated using preoperative routine blood tests. Receiver operating characteristic (ROC) curve was used to obtain the optimal cutoff value of SII. The Kaplan-Meier method was used to draw survival curves and survival rates were compared by log-rank test. The Cox proportional risk model was used to analyze single and multiple factors.Results:The SII area under the ROC curve in predicting postoperative survival was 0.749(95% CI: 0.641-0.858), the optimal threshold was 412.6. Using this threshold, patients were divided into the low SII group (SII≤412.6, n=80) and the high SII group (SII>412.6, n=101). The 1, 3, and 5-year cumulative survival rates of patients in the low SII group were 87.5%, 57.5%, and 26.3%, which were significantly better than those of the high SII group of 71.3%, 39.6%, and 9.9% respectively ( P<0.05). Multivariate analysis showed that SII>412.6 ( HR=2.887, 95% CI: 2.256-7.903, P<0.05) was an independent risk factor for overall survival of patients with hilar cholangiocarcinoma. Conclusion:Preoperative SII had predictive values for postoperative survival of patients with hilar cholangiocarcinoma, SII>412.6 was an independent risk factor for postoperative survival.

To study the chemical constituents of Cymbopogon citratus, isolation and purification of constituents were carried out on silica gel, Sephadex LH-20 and prepatative HPLC. The structures of the compounds were identified by physicchemical properties and spectral data analysis. Eight compounds were isolated and identified as 3beta-methoxy lanosta-9(11)-en-27-ol (1), 3beta-hydroxylanosta-9 (11)-en (2), (24S) -3beta-methoxylanosta-9(11), 25-dien-24-ol (3), 8-hydroxyl-neo-menthol (4), (2E)-3,7-dimethyl-2,7-octadiene-1, 6-diol (5), (+)-citronellol (6), 7-hydroxymenthol (7) and ethyl nonadecanoate(8). Compounds 1 is a new one. Compounds 2-3 are obtained from C. citratus for the first time.
Cymbopogon ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Triterpenes ; chemistry

OBJECTIVETo investigate the correlation between genotypes and biochemical phenotypes of phenylalanine hydroxylase (PAH) in patients with phenylketonuria (PKU).

METHODSThirteen exons and flanking introns of PAH gene in 102 patients with high blood phenylalanine levels (Phe > 120 umol/L) at initial diagnosis were amplified with polymerase chain reaction and analyzed with single strand conformation polymorphism (SSCP), denaturing high performance liquid chromatography (DHPLC) and DNA sequencing. Correlation between genotypes and biochemical phenotypes was analyzed.

RESULTSBiochemical assaying has indicated that 69 patients had classical PKU (Phe> 1200 umol/L), 31 were moderate (Phe 600-1200 umol/L), and 2 were mild (Phe 400-600 umol/L). More than 41 mutations and 75 genotypes have been identified. There were 9 (8.8%) homozygous mutations, which included 3 cases with R111X/R111X, 1 case with IVS4-1G>A/IVS4-1G>A, 3 cases with R243Q/R243Q and 2 cases with V399V/V399V. Among these 8 belonged to classic PKU phenotypes, except for a R243Q/R243Q genotype which has led to a moderate phenotype. In 91 patients carrying compound PAH mutations, 61 were classic, 29 were moderate, and 1 was mild. Patients who were heterozygous for R111X/R243Q and EX6-96A>G(Y204C)/R243Q were found with both classic and moderate PKU phenotypes. Certain individuals who have carried 2 null mutant alleles such as R111X/V399V, EX6-96A>G/Y356X and EX6-96A>G/V399V only showed a moderate phenotype. Individuals with R111X/A165D and R176X/A165D genotypes, on the other hand, respectively presented moderate and classic PKU phenotypes.

CONCLUSIONNinety percent of our patients are compound heterozygotes. Independent assortment of mutant alleles has resulted in a complex genotype-phenotype correlation. Although in most cases a correlation may be found, caution should still be taken upon genetic counseling. The phenomena where similar or even identical genotype may give rise to different biochemical phenotypes have implied that other factors may also influence the phenylalanine metabolism.

Adolescent ; Alleles ; Child ; Child, Preschool ; Exons ; Female ; Gene Frequency ; Genetic Association Studies ; Genotype ; Humans ; Infant ; Infant, Newborn ; Introns ; Male ; Mutation ; Phenotype ; Phenylalanine Hydroxylase ; genetics ; metabolism ; Phenylketonurias ; genetics ; metabolism

OBJECTIVETo observe the role of tumor necrosis factor α (TNF-α) in endothelial-mesenchymal transition (EnMT), and to explore the mechanism of fibrosis disease.

METHODSHuman umbilical vein endothelial cells (HUVEC) from umbilical cord of healthy fetus were isolated by enzymatic digestion and identified by immunofluorescence assay. The third to fifth generations of cultured HUVEC in logarithmic phase were harvested and seeded in 12-well plates and 6-well plates, and they were divided into control group (ordinary culture without any stimulation), 5, 10, 25, 50, and 100 ng/mL TNF-α groups (5, 10, 25, 50, 100 ng/mL of TNF-α was respectively added into the nutrient solution) according to the random number table, with three samples in each group. After being cultured for 72 hours, the cell morphology was observed under inverted phase-contrast microscope; the expression levels of coagulation factor VIII and α smooth muscle actin (α-SMA) were detected by immunofluorescence assay, and the ratios of numbers (absorbance values) of cells with expression of both factors were calculated. The mRNA expression levels of cadherin, α-SMA, and type I collagen were detected by RT-PCR (denoted as gray value ratio). Data were processed with one-way analysis of variance and LSD test.

RESULTS(1) The shape of primary HUVEC was round, short-spindle, or flat, and cells grew vigorously in cobblestone appearance after passages. After being subcultured for 1, 2, 3, 4, 5 passage (s), the positive rate of coagulation factor VIII of HUVEC was respectively (85.5 ± 1.8)%, (88.1 ± 5.0)%, (93.6 ± 3.7)%, (92.9 ± 4.8)%, (89.5 ± 1.1)%, and they were significantly higher than that of primary HUVEC [(81.4 ± 3.8)%, with F values all equal to 7.481, P values all below 0.05]. (2) As compared with that in control group, the appearance of cells in 5, 10, 25, 50, and 100 ng/mL TNF-α groups was gradually transformed from round, short-spindle, or flat shape to long-spindle shape with reduced intercellular junction and larger intercellular gap along with the increase in the concentration of TNF-α. (3) The ratios of numbers and the absorbance values of coagulation factor VIII and α-SMA double positive cells in control group (0.055 ± 0.015, 0.078 ± 0.017) were significantly lower than those in 5, 10, 25, 50, and 100 ng/mL TNF-α groups (0.257 ± 0.106, 0.280 ± 0.129, 0.505 ± 0.059, 0.817 ± 0.035, 0.929 ± 0.101 and 0.437 ± 0.040, 0.456 ± 0.097, 0.496 ± 0.082, 0.787 ± 0.131, 0.885 ± 0.087, with F value respectively 45.009, 50.099, P values all below 0.01). (4) The expression levels of cadherin mRNA in 5, 10, 25, 50, and 100 ng/mL TNF-α groups were 0.70 ± 0.05, 0.63 ± 0.06, 0.60 ± 0.10, 0.45 ± 0.16, and 0.26 ± 0.14, and it was significantly lower in the latter four groups than in control group (0.83 ± 0.03, with F values all equal to 11.593, P < 0.05 or P < 0.01). The mRNA expression levels of α-SMA and collagen I in 5, 10, 25, 50, and 100 ng/mL TNF-α groups were 0.45 ± 0.10, 0.51 ± 0.16, 0.49 ± 0.12, 0.60 ± 0.09, 0.76 ± 0.03 and 0.38 ± 0.18, 0.45 ± 0.15, 0.52 ± 0.12, 0.66 ± 0.17, 0.76 ± 0.20, and they were significantly higher in the latter three groups than in control group (0.37 ± 0.14, 0.31 ± 0.12, with F value respectively 7.839, 2.898, P < 0.05 or P < 0.01).

CONCLUSIONSTNF-α can obviously promote EnMT in a dose-dependent manner. EnMT may be another significant source of myofibroblasts that contributes to fibrotic tissue in scar formation.

Cell Differentiation ; Cells, Cultured ; Epithelial-Mesenchymal Transition ; drug effects ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Stromal Cells ; cytology ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVE: To study the moderated mediation for attention deficit/hyperactivity disorder (ADHD) with the symptoms of anxiety in children. METHODS: A total of 12 271 students were included with an average age of 8.9±1.9 years, including 6 743 male students and 5 508 female students, and 20 students with missing data on gender. Child psychological trauma questionnaires (parents version) and Conners questionnaires (parent version) were completed by the parents of primary school students. The data was studied by univariate analysis, multivariate analysis and moderated mediation analysis. RESULTS: The results of the univariate analysis showed that in all subjects, boys, and girls, the scores of hyperactivity index and childhood trauma were positively correlated with the score of anxiety (P<0.01), and ADHD and childhood trauma positively predicted anxiety disorder (P<0.001). The results of the multivariate analysis showed that in all subjects, boys, and girls, the scores of hyperactivity index (ADHD symptoms) and childhood trauma positively predicted the score of anxiety (P<0.001), and both ADHD and childhood trauma positively predicted anxiety disorder (P<0.001). The results of the moderated mediation analysis showed that childhood trauma was a mediating factor for the relationship between hyperactivity index and anxiety index in boys and girls (P<0.05), and sex moderated the relationship between hyperactivity index and anxiety index (P<0.001). CONCLUSIONS ADHD symptoms/ADHD are closely associated with anxiety symptoms/anxiety disorder. Childhood trauma exerts a mediating effect on the relationship between ADHD symptoms and anxiety symptoms, and sex moderates the relationship between ADHD symptoms and anxiety symptoms.
Anxiety ; Anxiety Disorders ; Attention Deficit Disorder with Hyperactivity ; Child ; Child Behavior Disorders ; Female ; Humans ; Male ; Surveys and Questionnaires

Objective To explore the effect of stromal cell-derived factor-1 α (SDF-1 α) on inducing recruitment of bone marrow-derived cells (BMDCs) to wound area. Methods BMDCs were isolated from bone marrow, cultured with routine method and identified by CXCR4 antibody. Cells cultured with CXCR4 antibody (100 ng,/mL) for 6 hours were labeled with CM-DiI and injected into the tail vein of full-thickness incisional wound model (set as anti-CXCR4 group). BMDCs labeled with CM-DiI without antibody treatment were injected to the rats in BMDCs group, and rats were injected with DMEM/F12 serum-free medium in the control group. The quantity of labeled BMDCs at the wound site and the percentage of wound closure were measured. Results (1) All BMDCs expressed CXCR4. (2) The percentages of wound closure at days 7 and 14 in BMDCs group (7 d: 41.3% ±4.6%; 14 d:92.3% ±2. 1%) were significantly higher than those of control group (7 d: 29.3% ±2. 3%; 14 d: 77.3% ±2.5%) and anti-CXCR4 group (7 d: 30.7% ±4.6% ;14 d: 85.7% ±1.5%) (P＜0.05). The percentage of wound closure of anti-CXCR4 group was significantly higher than that of control group at day 14(P ＜ 0.05). (3) The number of CM-DiI labeled BMDCs at wound site at days 7 and 14 in BMDCs group [7 d: (535 ±84) cells/hpf; 14 d: (769 ±124) cells/hpf) were greater than those of anti-CXCR4 group [7 d: (335 ±97) cells/hpf; 14 d: (521 ± 127) cells/hpf] (P＜0.05). Conclusions BMDCs participate in the cutaneous wound healing. SDF-1α plays an important role in recruiting BMDCs to wound area.