Esters ; urine ; Humans ; Phthalic Acids ; urine

11.0 for the pH of reaction solution,65℃～75℃for the reaction temperature,and 1:1.7 for the mass ratio of corn(dry weight)to FeCl_3?6H_2O.The corn polysaccharide-Fe(Ⅲ)complex synthesized with the optimized process was stable,with good solubility in water.The assay of Fe(Ⅲ)was 39.86%,40.20%and 40.17%,respectively for three batches of products.The RSD was

Background Retinopathy of prematurity is mainly due to retinal neovascularization.Objective This laboratory work was to evaluate the efficacy of different dosage of avastin for inhibiting retinal neovascularization.Methods Ninety 7-day-old clean C57BL/J6 mice were randomized into six groups as follows:air control group,hyperxia control group,hyperxia BSS group and avastin groups.C57BL/J6 mice in air control group were raised in regular air environments.The fifty mice were fed under the environment with 75％ ±2％ oxygen for 5 days to establish the retinal neovascularization models.The 1.25,2.50 and 5.00 g/L avastin (0.5 μl) were injected inteavtreally in forty-five mice models as low,moderate and high dosage avastin groups respectively,and 0.5 μl BSS was used at the same way in fifteen models as hyperxia BSS group.The mice were sacrificed in the 17-day-old age using excessive anesthesia method and the retina sections were prepared for the calculation of the numbers of vascular endothelial cell nuclei broken retinal inner membrane after hemotoxylin and eosin staining.The expression of CD34 in the retina was detected by immunochemistry.The morphology and distribution of retinal neovascular vessel in various groups were observed using retinal flat.The use of the animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The numbers of cell nuclei broken the inner limiting membrane was significant increased in the hyperxia group compared with the air control group( P＜0.01 ),and those in difference doses of avastin were considerably reduced in comparison with hyperxia BSS group (P＜0.01) and hyperxia group (P＜0.01 ).The decrease of numbers of cell nuclei broken the inner limiting membrane was obvious in low dose of high dose of avastin compared with low dose of avastin (P＜0.05 ).CD34 was positively expressed in retina internal membrane of hyperxia group.Retinal flat revealed the regular distribution and normal structure of retinal vessels in air control group and avastin groups.However,retinal and vitreous cavity neovascularization,leakage and enlarged non-perfusion regions in the perimeter of the retina were seen in hyperxia group and hyperxia BSS group. Conclusions Intravitreal injection of avastin can arrest retinal angiogenesis in oxygen-induced retinal neovascularization models in a dose-dependent manner.

Objective To investigate the role of T helper 17 cells/interleukin?17(Th17/IL?17) axis in the occurrence of vaginal candidiasis in mice. Methods A total of 120 female BALB/c mice were randomly and equally divided into Ei, En, Ci and Cn groups. Three days before vaginal inoculation, estrogen (Ei and En)groups and control(Ci and Cn)groups received subcutaneous injection of 0.05 mg estradiol and 0.1 ml sterilized soybean oil at the hind legs, respectively, and then the hormone treatment continued every other day until the end of experiment. Infected(Ei and Ci)groups and noninfected(En and Cn) groups were inoculated intravaginally with 10μl(5 × 104 conidia)of Candida albicans suspension and 10μl of sterilized phosphate?buffered saline, respectively. Ten mice were randomly selected from each group and sacrificed on day 3, 7 and 14 after inoculation. The intact vagina tissues were resected and then frozen in liquid nitrogen or embedded in paraffin. Real?time fluorescence?based quantitative PCR(qRT?PCR)and immunofluorescent staining were performed to measure mRNA expression and immunofluorescence intensities of retinoic acid?related orphan receptorγt(RORγt), RORα and IL?17, respectively. Western blot analysis was conducted to determine protein expression of RORγt and IL?17. Results Laser scanning confocal microscopy showed that RORγt, RORα and IL?17 immunofluorescence was mainly located at inflammatory cells of the lamina propria and blood vessels in En and Cn groups, at mucosal epithelium, adherent hyphae, and inflammatory cells of the lamina propria and blood vessels in Ci group, and at mucosal epithelium, vaginal canal and endocytosed hyphae, and inflammatory cells of the lamina propria and blood vessels in Ei group. qRT?PCR and immunofluorescent staining uncovered that mRNA expression and immunofluorescence intensities of RORγt, RORα and IL?17 were significantly higher in En, Ci and Ei groups than in Cn group at the same time points(all P<0.05), as well as in the Ei group than in En and Ci groups(both P<0.05), and were increased gradually over time in En, Ci and Ei groups, but not in the Cn group. Additionally, mRNA expression and immunofluorescence intensities of RORγt and RORαand IL?17 generally peaked on day 14 after inoculation, while the immunofluorescence intensity of IL?17 peaked on day 7 (P < 0.05). Western blot analysis revealed that protein expression of RORγt and IL?17 was significantly higher in the infected(Ei and Ci)groups than in the noninfected(En and Cn)groups at the same time points(RORγt:F=45.685, P<0.001;IL?17:F=29.655, P<0.01), and was highest in the Ei group(P<0.05);however, no significant differences were observed between Cn and En groups(both P>0.05). Moreover, RORγt and IL?17 protein expression in Ci and Ei groups was obviously up?regulated on day 7 after inoculation (RORγt: F = 13.137, P < 0.001; IL?17: F = 11.182, P < 0.001), but was not increased further on day 14. Conclusion Vaginal candida infection can up?regulate the expression of RORγt, RORα and IL?17, suggesting that Th17/IL?17 axis may be involved in the occurrence of vaginal candidiasis in BALB/c mice.

Objective The microglias is the representative of immune cells in the brain. It plays dual roles of both repairing and damaging in injured nervous system, and works as an inevitable component of the circumstance of injured neurons. This study was aiming at the effects of the microglias on the biological activities of mesenchymal stem cells (MSCs) in the circumstance of injured neurons. Methods MSCs were obtained by primary culture. We adopted PC12 cells (PC12) and BV2 cells (BV2) to substitute for neurons and microglias, respectively. PC12 were injured by aged Abeta(1-40) and the supernatant of the injured PC12 was used to set up the circumstance of injured neurons. Transwells were used for co-culture of BV2 and MSCs, which allowed the independent detection of cells after co-culture. Immunofluorescence was used to identify MSCs and neuron-differentiating cells with CD44 and neuron specific enolase (NSE) staining, respectively. MTT assay was adopted to measure the proliferation. Results In the circumstance of both BV2 presence and injured PC12 supernatant incubation, either the proliferation or the differentiation of MSCs reached the highest, which seemed to be contradictory, but we gave our explanations. With the BV2 co-culture, the proliferation of MSCs tend to be higher, but the neuron-differentiating MSCs were similar to those incubated without BV2 co-culture either in normal or injured in PC12 supernatant. With the incubation of injured PC12 supernatant, the neuron-differentiating cells were significantly higher than that of control (P < 0.05). Conclusion In the circumstance of injured neurons, microlgias tend to promote the MSCs proliferation. Although not helpful in neuron-differentiating, microglias did not exert any negative effect either.

Objective To investigate the role of H(1) and H(2) receptors in the locus ceruleus (LC) in carotid sinus baroreceptor reflex (CSR) resetting induced by intracerebroventricular (i.c.v.) injection of histamine (HA). Methods The left and right carotid sinus regions were isolated from the systemic circulation in 18 male Sprague-Dawley rats anesthetized with pentobarbital sodium. The intracarotid sinus pressure (ISP) was altered in a stepwise manner in vivo. ISP-mean arterial pressure (MAP) relationship curve and its characteristic parameters were constructed by fitting to the logistic function with five parameters. The changes in CSR performance induced by i.c.v. HA and the effects of pretreatment with H(1) or H(2) receptors selective antagonist, chlorpheniramine (CHL) or cimetidine (CIM) into the LC, on the responses of CSR to HA were examined. Results I.c.v. HA (100 ng in 5 mu l) significantly shifted the ISP-MAP relationship curve upwards (P < 0.05) and obviously decreased the value of the reflex parameters such as MAP range and maximum gain (P < 0.05), but increased the threshold pressure, saturation pressure and ISP at maximum gain (P < 0.05). The pretreatment with CHL (0.5 mu g in 1 mu l) or CIM (1.5 mu g in 1 mu l) into the LC could obviously attenuate the changes mentioned above in CSR performance induced by HA, but the alleviative effect of CIM was less remarkable than that of CHL (P < 0.05). Respective microinjection of CHL or CIM alone into the LC with the corresponding dose and volume did not change CSR performance significantly (P > 0.05). Conclusion Intracerebroventricular administration of HA results in a rapid resetting of CSR and a decrease in reflex sensitivity, and the responses of CSR to HA may be mediated, at least in part, by H(1) and H(2) receptors activities in the LC, especially by H(1) receptors. Moreover, the effects of the central HA on CSR might be related to a histaminergic descending pathway from the hypothalamus to LC.

OBJECTIVETo study the effect of occupational stress on cardiovascular function of different vocational population.

METHODSThe occupational stressors, risk factors of cardiovascular diseases were investigated by questionnaire in 839 people with 4 kinds of jobs. Blood pressure, sugar, and lipid were detected at the same time.

RESULTSBlood pressure were higher in the groups of old age, long standing and teachers, and the abnormal rate of blood pressure was 21.69%. There was no difference in abnormal ECG among ages, standing and occupation, and the abnormal rate of ECG was 19.07%. Job control, job demands, job responsibility, role in a job and shift work were the main stress factors affecting systolic and diastolic blood pressure. More conflict in job, less chance of participation, severe job loads were the risk factors of primary hypertension. Accident due to job responsibility, job responsibility, role in a job were the main risk factors of abnormal electrocardiograph. Self-respect and activity beyond work were the good modifiers of heart function.

CONCLUSIONOccupational stress has certain effect on cardiovascular function.

Adult ; Blood Pressure ; Electrocardiography ; Female ; Humans ; Logistic Models ; Male ; Occupational Diseases ; physiopathology ; Stress, Psychological ; physiopathology

OBJECTIVETo screen the genes related with leukocyte responses in mice early after burn injury by bioinformatic analysis of the gene expression profiling data.

METHODSGene expression profiles were obtained from GEO (GSE7404, Mouse musculus, 25% TBSA, full-thickness) database. After screening of the differentially expressed genes (DEGs) through paired-sample t-test and fold-change, DAVID online tools were used to select the DEGs related to leukocyte responses to burns by GO functional enrichment analysis; the interacting genes identified through KEGG pathway enrichment analysis were transferred to STRING to construct the protein-protein interaction (PPI) network. Biological annotation of the sub-networks was executed using the software Cytoscape. Real-time PCR was used to verify the DEGs identified in mice.

RESULTSOf the 259 leukocyte response-related DEGs screened at 1 day post-burn, 118 were up-regulated and 141 were down-regulated. KEGG pathway enrichment analysis showed that the pathways were associated with the immune function, cell growth and cell death. PPI network and module analysis suggested that some of genes (such as Lck, Stat1, Myd88, Stat3, and Jun) play critical roles in the PPI network post-burn. RT-PCR results were consistent with those of bioinformatic analysis.

CONCLUSIONSLck, Stat1, Myd88, Stat3, and Jun might be critical players in the development of leukocyte response in mice early after burn injury. Our finding provides new insights into the pathogenesis of leukocyte response to burn injury and identifies several potential biomarkers for burn treatment.

Animals ; Burns ; genetics ; Computational Biology ; Down-Regulation ; Gene Expression Profiling ; Gene Regulatory Networks ; Leukocytes ; physiology ; Mice ; Real-Time Polymerase Chain Reaction ; Software ; Up-Regulation

OBJECTIVETo study the effect of low- and intermediate-dose factor VIII (FVIII) for prophylactic treatment of severe hemophilia A in children by comprehensively evaluating the outcomes of the joints.

METHODSForty-seven children with severe hemophilia A (FVIII activity ≤2%) were enrolled in this study. Eighteen of the children received prophylactic treatment with low-dose FVIII (10 U/kg, 2-3 times a week), 20 received prophylactic treatment with intermediate-dose FVIII (15-30 U/kg, 3 times a week), and 9 received on-demand treatment with FVIII infusion when bleeding occurred according to the Chinese Expert Consensus on the Diagnosis and Treatment of Hemophilia. The children were followed up for 180 days to observe the changes in the indexes of clinical bleeding phenotype, joint structure, joint function, and joint mobility, and the correlation of these indexes were analyzed.

RESULTSCompared with on-demand treatment, prophylactic treatment with low- and intermediate-dose FVIII significantly improved the clinical hemorrhage phenotype (P<0.01), and the improvement was significantly more conspicuous with intermediate-dose prophylactic treatment (P<0.05). Comprehensive evaluation of the joint structure and function changes showed that compared with on-demand treatment, prophylactic treatment with low- and intermediate-dose FVIII resulted in significant improvements in the total score of Hemophilia Joint Health Score (HJHS), Functional Independence Score in Hemophilia (FISH), the single most severe target joint ultrasound and HJHS score of the target joint (P<0.05) and prophylactic treatment with intermediate-dose FVIII appeared to produce better outcomes of the joint than low-dose FVIII. No correlation was found between annual target joint bleeding rate (ATJBR) and ultrasound score, between ATJBR and HJHS change, or between annual joint bleeding rate (AJBR) and the total score of FISH (P>0.05).

CONCLUSIONCompared with on-demand treatment, prophylactic treatment with low- and intermediate-dose FVIII can significantly improve the bleeding phenotype and delay the progression of joint injury, but the clinical hemorrhagic phenotype is not sufficient to monitor the disease progression.

Objective:To investigate the role of folliculin in apoptosis of and chemokine secretion by melanocytes mediated by interferon-γ (IFN-γ) .Methods:Normal primary melanocytes were isolated from circumcised foreskin tissues from a healthy male child, and primary vitiliginous melanocytes were isolated from normally pigmented suction-blistered epidermis from patients with vitiligo after suction blister epidermal grafting. Western blot analysis was performed to determine the folliculin protein expression in normal primary melanocytes, primary vitiliginous melanocytes and a human primary melanocyte line PIG1. PIG1 cells stimulated with 10 ng/ml IFN-γ for 48 hours served as induction group, and untreated PIG1 cells served as control group. Real-time quantitative RCR (qRT-PCR) was performed to determine the mRNA expression of folliculin, autophagy-related microtubule-associated protein 1 light chain 3 (LC3) -Ⅱ and Beclin genes, and Western blot analysis to determine the protein expression of folliculin, Beclin1 and LC3Ⅱ/Ⅰ, as well as phosphorylation levels of adenosine monophosphate-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) in the above cells. Furthermore, the melanocytes stimulated with 10 ng/ml IFN-γ for 48 hours were divided into several groups: negative control group infected with an empty lentiviral vector, folliculin inhibition group infected with a folliculin-inhibiting lentivirus, autophagy enhancement group infected with a folliculin-inhibiting lentivirus followed by 2-hour treatment with a mTOR inhibitor, autophagy inhibition group infected with a folliculin-inhibiting lentivirus followed by 2-hour treatment with an AMPK inhibitor. Then, flow cytometry was conducted to detect apoptosis of PIG1 cells, and enzyme-linked immunosorbent assay to measure the concentration of chemokines CXCL10 and CCL20 in the culture supernatant of PIG1 cells in the above groups. Measurement data were compared among multiple groups by using one-way analysis of variance, and multiple comparisons were carried out by using least significant difference- t test. Results:The relative protein expression level of folliculin significantly differed among the normal primary melanocytes (0.850 ± 0.120) , primary vitiliginous melanocytes (1.507 ± 0.170) and PIG1 cells (0.697 ± 0.130; F = 50.09, P < 0.001) , and was significantly higher in the primary vitiliginous melanocytes than in the normal primary melanocytes and PIG1 cells ( t = 4.06, 5.89, respectively, both P < 0.01) . Compared with the control group, the induction group showed significantly increased relative mRNA and protein expression levels of folliculin (both P < 0.01) , but significantly decreased relative mRNA and protein expression levels of LC3Ⅱ and Beclin (all P < 0.01) ; moreover, the induction group showed significantly decreased LC3Ⅱ/Ⅰ levels (0.72 ± 0.02) and AMPK phosphorylation levels (0.714 ± 0.023) in the PIG1 cells compared with the control group (1.13 ± 0.02, 1.176 ± 0.002, t = 7.34, 6.67, respectively, both P < 0.01) , but significantly increased mTOR phosphorylation levels (1.051 ± 0.023) compared with the control Group (0.451 ± 0.016, t = 3.81, P = 0.009) . There were significant differences in the PIG1 cell apoptosis rate and concentrations of CXCL10 and CCL20 among the control group, induction group and other treatment groups (all P < 0.001) ; specifically, the PIG1 cell apoptosis rate and concentrations of CXCL10 and CCL20 were significantly higher in the induction group than in the control group, lower in the folliculin inhibition group than in the negative control group, lower in the autophagy enhancement group than in the folliculin inhibition group, and higher in the autophagy inhibition group than in the folliculin inhibition group (all P < 0.05) . Conclusions:Folliculin is highly expressed in vitiliginous melanocytes. Folliculin expression and downstream signaling pathways are regulated by IFN-γ, and folliculin may participate in IFN-γ-mediated melanocyte apoptosis and chemokine secretion via regulating autophagy.