Objective · To investigate effect of fucoidan on autophagy, migration and invasion in human multiple myeloma U266 cells. Methods · The U266 cells treated with fucoidan were cultured in vitro. The formation of autophagosomes was observed by transmission electron microscopy (TEM). Transwell assay was used to evaluate the effect of fucoidan on migratory and invasive abilities of U266 cells. The protein levels of LC3-Ⅱ/LC3-Ⅰ, Beclin-1, P62, MMP9, CXCR4, p-AKT/T-AKT, p-mTOR/T-mTOR were detected by Western blotting. MMP9 concentration in the culture medium was examined by ELISA. Results · ① Autophagosomes increased in fucoidan-treated cells compared with control group under TEM. ② Migratory and invasive abilities were inhibited by fucoidan in a dose-dependent manner, which were suppressed by chloroquine. ③ Western blotting demonstrated that expression of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and MMP9 increased in fucoidan-treated cells, while P62, CXCR4, p-AKT/T-AKT and p-mTOR/T-mTOR decreased compared with control group. ④ The result of ELISA showed that MMP9 concentration in the culture medium of fucoidan-treated cells significantly decreased. Conclusion · Fucoidan induces autophagy and inhibits migration and invasion in U266 cells.

Objective] To evaluate effect of health literacy intervention in occupational groups , explore suitable ways and provide suggestions for workplace health promotion . [ Methods ] In application of convenience sampling , Minhang District , Hongkou District and Qingpu District each selected 2 enterprises respectively according to their type , scale and location , setting one as intervention enterprise and the other as control .Then a six-month comprehensive intervention was conducted among intervention groups .Two hundred employees in each enterprise were randomly investigated before and after intervention to evaluate the effect . [ Results ] The overall health literacy level of participants in the intervention group rose from 16.7% to 33.4%, with improvement on the literacy levels of scientific view , infectious diseases prevention and treatment , chronic disease control and prevention and the three aspects of health literacy content ( P<0 .05 ) , while such improvement was not reflected in the control group , and health skill literacy, safety and first aid literacy declined (P <0.05). [Conclusion] Comprehensive intervention based on health needs could improve the level of health literacy of occupational groups . Intervention in future should focus on improving the literacy of chronic disease control and prevention and basic medical care in occupational population .

OBJECTIVETo study the incidence of various enzyme deficiency in tetrahydrobiopterin (BH4) metabolism and the related gene mutation among the patients with motor disturbance and mental retardation.

METHODSOne hundred patients with unknown motor disturbance and mental retardation were referred to this study. All patients were performed by phenylalanine (Phe) and BH4 loading test, urinary pterin analysis and dihydropteridine reductase (DHPR) activity. Some patients received the dopa treatment for diagnosis of dopa-responsive dystonia (DRD). The analysis of GTP cyclohydrolase 1 gene (GCH1) mutation for DRD patients and the analysis of 6-pyruvoyl tetrahydropterin synthase (PTS) gene mutations for PTS deficient patients were done under the consent from their parents.

RESULTSSeventy of 100 patients had normal basic blood Phe levels, six (6%) patients were diagnosed as DRD. Thirty patients had hyperphenylalaninemia (HPA), eight (8%) were diagnosed as PTS deficiency and 22(22%) were diagnosed as phenylalanine hydroxylase (PAH) deficiency. All patients had normal DHPR activity. The mutation IVS5+3insT of GCH1 was found in 2 patients with DRD. Seven kinds of PTS mutations were found in 8 patients with PTS deficiency, and 75% of the mutations were 259C-->T,286G-->A and 155A-->G.

CONCLUSIONSome patients with unknown motor disturbance and mental retardation may suffer from BH4 metabolism related diseases. Theses patients are necessary to be screened for such kind of diseases in order to confirm the diagnosis.

Adolescent ; Biopterin ; analogs & derivatives ; metabolism ; Child ; Child, Preschool ; Dihydropteridine Reductase ; genetics ; metabolism ; Dystonia ; genetics ; metabolism ; Female ; GTP Cyclohydrolase ; genetics ; metabolism ; Humans ; Infant ; Intellectual Disability ; genetics ; metabolism ; Male ; Mutation ; Phenylalanine Hydroxylase ; genetics ; metabolism ; Phosphorus-Oxygen Lyases ; genetics ; metabolism

Objective To investigate whether the protective effects of leonurine (SCM-198) against endotoxin induced uveitis (EIU) of SD rats caused by lipopolysaccharide (LPS) was existing,and discuss the underlying mechanisms.Methods Thirty-six normal healthy male SD rats were divided into 3 groups randomly with the same baseline bodyweight and feeding conditions.All rats received intragastric administration every day.The experimental group was devided into 4 subgroups,rats in these subgroups received SCM-198 intragastric administration by as the dose of 10,20,40 and 80 mg/kg bodyweight per day,rats in the negative control group received intragastric administration of normal saline 10 mL/kg per day,rats in the positive control group received intragastric administration ofdexamethasone (DEX) 0.5 mg/kg bodyweight per day.All rats received a 21-day-intragastric administration.The body weight of all rats was monitored every 7 days.The electroretinogram (ERG) examination was taken in the 18th day.All rats received a 100 mg S.typhi LPS intraperitoneal injection after the 21st intragastric administration.Twenty-four hours later,following anaesthesia,all rats received another ERG examination,and inflammation was scoring under microscope by 2 experienced ophthalmologists,after that the aqueous humor of all rats was collected from the left eye.The aqueous humor was kept in-80 ℃ immediately.Then the rats were sacrificed and the right eyes were immediately enucleated to finish the HE staining and immunohistochemistry (IHC) staining examination of tumor necrosis factor-α (TNF-α) and intercellular cell adhesion molecule-1 (ICAM-1).The total amount of protein in aqueous humor was detected by BCA test.Western blot was used to examine the expression of TNF-α,interleukin-1β (IL-1β),IL-6 and ICAM-1.All data was analyzed by SPSS 19.0,and differences were considered significant at P＜0.05.Results The body weight of the rats in positive control group was significantly lower (P＜0.05) than the experimental group and the negative control group after the 21-day-intragastric administration.The inflammatory score of experimental group was lower than that of the negative control group,but higher than the score of positive control group.The HE staining sections showed the similar results.The a wave of ERG in 0.01 cd of rats received 20 mg/kg SCM-198 daily intragastric administration after LPS injection was significantly lower than that before the LPS injection (P＜0.05),also lower than other groups after LPS injection.The expression of TNF-α,IL-6 and IL-1β in the aqueous humor of the rats in the subgroup of SCM-198 10 mg/kg daily intragastric administration was lower than other groups.Conclusions Intragastric administration of SCM-198 has protective effect against endotoxin induced uveitis in SD rats without obvious adverse reaction,which could alleviate the imflammatory reaction and the damage to the uvea construction.NF-κB plays an important role in the reaction.Thus,SCM-198 is a candidate potent compound with potential therapeutic applications in inflammation associated eye diseases.While the best mode and dose of administration should be further investigated.

OBJECTIVETo determine distribution and mutation pattern of type P ATP7B gene mutation and to explore genotype and phenotype correlation in patients with Wilson's disease (WD).

METHODSSixty patients with WD from 57 no kinship families, 37 male and 23 female, were enrolled in this study. The age of onset ranged from 4.6 - 39 years, < or = 16 years in 55 patients. Some exons of ATP7B gene mutation were analyzed in patients with WD by using biochemical methods, polymerase chain reaction-single strand configuration polymorphism (PCR-SSCP), restriction fragment and DNA sequence analysis. Totally 778 coding regions were identified with restriction enzyme Msp I. The activity of Cu-ATPase was assessed by measuring inorganic phosphorus in 3 patients with known genotype.

RESULTSFifty-two of 60 patients (86%) had presented with hepatic manifestations, 30 of them had only hepatic manifestations, 12/52 patients had hepatic and neurological manifestations at the same time; 10/52 patients had hepatic and other symptom; 7/60 patients had only neurological symptom, one patient had no symptom. Eleven mutations were detected by DNA sequencing, including five missense mutations (R778L, V1140A,G943S, V1106I and V1216M), one deletion (1384del17) and five polymorphisms (IVS4-5T/C, A2495G, C2310G, IVS18 + 6C/T and IVS20 + 5A/G) were identified. R778L mutation was identified 52/114 alleles (45.6%). R778L occurred in 38/52 patients with hepatic manifestation (73%), homozygosis of R778L was demonstrated in 14 patients and heterozygosity of R778L in 24 patients. V1106I mutation was 1.7%, G943S, V1140A, and V1216M was 0.86% respectively in this study. Two patients with delayed onset of neurological symptoms occurred V1106I mutation of ATP7B. Cu-ATPase activity of 3 patients with known mutation (R778L/V1106I, R778L/V1216M and R778L/R778L) declined by 44.55%, 88.23% and 69.49%, respectively, compared with normal control.

CONCLUSIONThe 1384del17bp and V1106I are two novel mutations found in patients with WD. R778L was common mutation of ATP7B gene with frequency of 45.6% in this study. The mutation in exon 8 of WD gene may play an important role in pathogenesis of WD in Chinese. Carriage of R778L mutation seems to be correlated with hepatic manifestation.

Adenosine Triphosphatases ; genetics ; Adolescent ; Adult ; Cation Transport Proteins ; genetics ; Child ; Child, Preschool ; Copper-transporting ATPases ; DNA Mutational Analysis ; Exons ; Female ; Gene Frequency ; Genotype ; Hepatolenticular Degeneration ; enzymology ; genetics ; pathology ; Humans ; Male ; Mutation ; Phenotype ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA

OBJECTIVETo confirm the diagnosis of multiple carboxylase deficiency (MCD) on the gene level and explore the mutations in Chinese children with MCD.

METHODSBiotinidase (BT) and holocarboxylase synthetase (HLCS) genes were analyzed by PCR and direct sequencing for the 4 BT deficiency patients and 8 HLCS deficiency patients, respectively. The identified mutations in the parents of the patients and 50 normal controls were screened by PCR-restriction fragment length polymorphism and direct DNA sequencing.

RESULTSTotal detection rate of gene mutation is 100% in the 12 children with MCD. Six mutations were detected in the 4 children with BT deficiency, they were c. 98-104del7ins3, c. 1369G>A (V457M), c. 1157G>A(W386X), c. 1284C>A(Y428X), c. 1384delA and c. 1493_1494insT. The last four were novel mutations. Four mutations were found in the 8 children with HLCS deficiency. They were c. 126G>T (E42D), c. 1994G>C (R665P), c. 1088T>A (V363D) and c. 1522C>T (R508W). The last two were hot-spot mutations [75%(12/16)], and c. 1994G>C (R665P) was a novel mutation.

CONCLUSIONThis study confirmed the diagnosis of 12 patients with MCD on the gene level. Six mutations were found in the BT gene and 4 in the HLCS gene, including 5 novel mutations. Two mutations of the HLCS gene are probably hot-spot mutations in Chinese children with HLCS deficiency.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Biotinidase ; genetics ; Biotinidase Deficiency ; Carbon-Nitrogen Ligases ; deficiency ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; Multiple Carboxylase Deficiency ; genetics ; metabolism ; Mutation

OBJECTIVETo study the clinical and laboratory characteristics of chronic active Epstein-Barr virus (EBV) infection (CAEBV) in children and to provide a basis for the diagnosis and treatment of CAEBV.

METHODSThe clinical data of 13 children with CAEBV, as well as 15 cases of acute EBV infection (AEBV) as controls, were analyzed, including clinical manifestations, EBV antibodies, EBV DNA, and peripheral blood lymphocyte subsets.

RESULTSBoth groups of patients had infectious mononucleosis-like symptoms such as fever, hepatomegaly, splenomegaly, and lymphadenectasis, but CAEBV patients had a longer course of disease and continuous and recurrent symptoms. Compared with the AEBV group, the CAEBV group had a significantly higher EBV DNA load in peripheral blood (P<0.05), a significantly higher VCA-IgG titer (P<0.05), and significantly lower numbers of white blood cells, lymphocytes, B cells, total T cells, CD4+ T cells, and CD8+ T cells in peripheral blood (P<0.05). Among 13 CAEBV patients followed up, 8 cases died, 2 cases showed an improvement, 2 cases had a recurrence, and 1 case was lost to follow-up after being transferred to another hospital. All the AEBV patients were cured and had no recurrence during the one-year follow-up.

CONCLUSIONSThe clinical manifestations of CAEBV vary in children. It is difficult to distinguish CAEBV from AEBV early. More attention should be paid to CAEBV because of its severe complications, poor prognosis, and high mortality. Measurement of EBV DNA load, VCA-IgG titer, and lymphocyte subsets in peripheral blood may be helpful in the diagnosis and differential diagnosis of CAEBV.

Adolescent ; Child ; Child, Preschool ; Chronic Disease ; Epstein-Barr Virus Infections ; diagnosis ; immunology ; virology ; Female ; Humans ; Infant ; Lymphocyte Subsets ; immunology ; Male

The antisense approach and RT-PCR were used to study the effects of muscarinic receptors on the scores of morphine-withdrawal syndrome and the expression of NMDA receptor subtypes (NR(1A) and NR(2A)) mRNA in rat spinal cord and brainstem. The concentrations of glutamate in periaqueductal grey (PAG) of morphine-withdrawal rats were determined by capillary electrophoresis with laser-induced fluorescence detection. The data showed that the NR(1A) and NR(2A) mRNA levels were increased significantly in the spinal cord and brainstem 1 h after the injection of naloxone (4 mg/kg, i.p.) in morphine-dependent rats. Moreover, in morphine-dependent rats pretreated (i.p.) with scopolamine (0.5 mg/kg), or pirenzepine (10 mg/kg), MK801 (0.125 mg/kg), L-N-nitroarginine methylester (10 mg/kg) 30 min before naloxone injection, the NR(1A) and NR(2A) mRNA levels were significantly lower than those of 1 h morphine-withdrawal rats. Intrathecal injection of NR(1A) or M(2) receptor antisense oligonucleotides (A-oligo, 4 microg/per rat) 24 h prior to naloxone challenge could block the morphine withdrawal symptoms including wet dog shaking, irritability, salivation, diarrhea, chewing and weight loss. Meanwhile, in morphine-dependent rats the NR(1A) mRNA levels in the spinal cord and brainstem were down-regulated by intrathecal injection of M(2) receptor A-oligo. The glutamate concentrations in PAG microdialysis were increased to a maximal level 15 min after naloxone injection. The glutamate response was inhibited by pretreatment with M(2) receptor A-oligo but not by M(1) A-oligo. The results suggest that the expression of NMDA receptors and the release of glutamate in brainstem are involved in the processes of morphine withdrawal and that the NMDA receptor expression is possibly regulated by the muscarinic receptors during morphine withdrawal.
Animals ; Brain Stem ; metabolism ; Glutamic Acid ; metabolism ; Male ; Morphine ; adverse effects ; Periaqueductal Gray ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Muscarinic ; physiology ; Receptors, N-Methyl-D-Aspartate ; biosynthesis ; genetics ; Spinal Cord ; metabolism ; Substance Withdrawal Syndrome ; genetics ; metabolism

AIMTo observe mRNA expression of muscarinic acetylcholine receptors in spinal cord and brainstem in morphine dependent or withdrawal rats.

METHODSThe mRNA expression level of m1, m2, m3, m4 and m5 were determined by RT-PCR, the beta-actin mRNA expression was used as internal control.

RESULTSThe mRNA level of m1, m2, m3, m4 and m5 in spinal cord and m1 and m2 in brainstem were increased significantly during morphine dependence, and the levels of m1, m2, m3 and m4 in spinal cord and m1 in brainstem were decreased 1 h after the injection of naloxone (4 mg.kg-1, i.p.) in morphine dependent rats. Either scopolamine (0.5 mg.kg-1) or pirenzepine (10 mg.kg-1) was shown to significantly decrease the morphine withdrawal symptoms in rats. The levels of m1, m2, m3 and m5 in spinal cord were increased by pretreatment with pirenzepine and the levels of m2, m3 and m4 in spinal cord were increased by pretreatment with scopolamine.

CONCLUSIONThe adaptive expression of muscarinic receptors at spinal and supraspinal levels play important role in mediating morphine dependence and withdrawal in rats.

Animals ; Brain Stem ; drug effects ; metabolism ; Gene Expression ; drug effects ; Male ; Morphine ; toxicity ; Morphine Dependence ; metabolism ; RNA, Messenger ; biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Receptors, Muscarinic ; biosynthesis ; classification ; genetics ; Spinal Cord ; drug effects ; metabolism ; Substance Withdrawal Syndrome ; metabolism

Column chromatographies over silica gel, Sephadex LH-20, reverse phase C18, and MCI, and semi-preparative HPLC were used for separation and purification of constituents from Inula cappa. The 22 compounds were obtained and their strutures were determined by NMR and MS spectra data as nine flavonoids: luteolin (1), apigenin (2), chrysoeriol (3), artemetin (4), 2', 5-di- hydroxy-3, 6, 7, 4', 5'-pentamethoxyflavone (5), chrysosplenol C (6), apigenin-5-0-β-D-glucopyranoside (7), luteolin-3-methyl, luteolin-3-methylether-4'-0-β-D-glucopyranoside (8), luteolin-4'-0-β-D-glucopyranoside (9); four triterpenes: darma-20, 24-dien- 3β-0-acetate (10), darma-20, 24-dien-3β-ol (11), epirfiedelanol (12), friedelin (13); three coumarins: scopoletin (14) , isosco- poletin (15) , scopolin(16) , and other types of compounds stigmasta-5, 22-dien-3β-0-7-one (17), stigmasterol (18), palmitic acid (19), linoleic acid (20), linoleic acid methyl ester (21), (E) -9, 12, 13-trihydroxyoetadee-10-enoie acid (22). Compound 5 is a new natural product. Compounds 3-9, 15, 17, 21, and 22 were isolated from this genus for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Inula ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization