Adult ; Aged ; Brucellosis ; diagnosis ; therapy ; Female ; Humans ; Male ; Middle Aged ; Occupational Diseases ; diagnosis ; microbiology ; therapy

Chickenpox ; complications ; Child ; Hemiplegia ; diagnosis ; virology ; Humans

Chemical investigation of fruits of Mours alba L. lead to the isolation of fifteen compounds by various chromatographies such as silica gel, Sephadex LH-20, RP-C18 column chromatography. Their structures were determined to be: 1-[5-(2-formlfuryl) methyl] dihydrogen 2-hydroxypropane-1, 2, 3-tricarboxylate 2, 3-diethyl ester (1), 1-[2-(furan-2-yl)-2-oxoethyl] pyrrolidin-2-one (2), divaricataester A (3), methyl 1-[2-(furan-2-yl)-2-oxoethyl]-5-oxopyrrolidine-2-carboxylate (4), 1-[2-(furan-2-yl)-2-oxoethyl]-5-oxopyrrolidine-2-carboxylic acid (5), L-pyroglutamic acid (6), L-pyroglutamic acid ethyl ester (7), 3-O-caffeoylquinic acid methyl ester (8), 3-O-caffeoylquinic acid ethyl ester (9), 5-O-caffeoylquinic acid methyl ester (10), 5-O-caffeoylquinic acid ethyl ester (11), 4-O-caffeoylquinic acid methyl ester (12), 4-O-caffeoylquinic acid methyl ester (13), 4-O-caffeoylquinic acid (14), 3-O-caffeoylquinic acid (15), respectively, based on the spectral analysis such as NMR, MS etc. Compounds 1-14 were isolated from this genus for the first time, among which 1 was a new compound.
Chlorogenic Acid ; isolation & purification ; Esters ; Fruit ; chemistry ; Furans ; chemistry ; isolation & purification ; Lactams ; isolation & purification ; Molecular Structure ; Morus ; chemistry ; Plants, Medicinal ; chemistry ; Pyrrolidonecarboxylic Acid ; isolation & purification ; Tricarboxylic Acids ; chemistry ; isolation & purification

This study is to investigate the effect of the effective components group of Xiaoshuantongluo (XECG) on neuronal injury induced by oxygen-glucose deprivation (OGD) in primary cortical cultures isolated from SD rat cortex at day 3 and the possible mechanism. Cells were divided into control group, OGD model group and XECG group (1, 3 and 10 mg x L(-1)). The cell viability was assessed with MTT assay and the LDH release rate was measured by enzyme label kit. The cell apoptosis was analyzed using Hoechst staining. RT-PCR was applied to detect the mRNA levels of JAK2 and STAT3. Western blotting was used to detect the expressions of Bcl-2, Bax, p-JAK2 and p-STAT3 proteins. Results showed that XECG resulted in an obvious resistance to oxygen-glucose deprivation-induced cell apoptosis and decrement of cell viability, decrease the cell LDH release rate. XECG could adjust the expression of Bcl-2 and Bax proteins and increase Bcl-2/Bax ratio, up-regulate the expression of p-JAK2 and p-STAT3. In conclusion, XECG could protect against the neuronal injury cells exposed to OGD, which may be relevant to the promotion of JAK2/STAT3 signaling pathway, and impact the expression of Bax and Bcl-2.
Animals ; Apoptosis ; Cell Survival ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; Janus Kinase 2 ; metabolism ; Neurons ; drug effects ; metabolism ; Neuroprotective Agents ; pharmacology ; Oxygen ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo prepare anti-recombinant protein antibody from immunized mice with recombinant nucleocapsid protein (NP) of human influenza A3 (IFV-A3) virus expressed in prokaryotic cell, and to explore the feasibility of utilizing anti-recombinant protein antibody to detect influenza A virus.

METHODSNP genes of human influenza A virus were analyzed with computer softwares of ClustalX, Antheprot, et al. to determine the antigenicity in conserved regions. Three different partial NP genes were harvested and cloned into pET-28(c) plasmid, the recombinant plasmids were induced to express partial NP segments in BL21 cells. The recombinant proteins were purified with Ni-agarose by affinity chromatography and immunized BALB/c mice. The polyclonal antisera harvested from mice were analyzed with Western Blot and immunohistochemistry assays to detect the reactions with IFV-A.

RESULTSThree recombinant plasmids were expressed with high yield in BL21 cells, about 15-20 mg/L. Western Blot results indicated that the three prepared antisera (1:2000) positively reacted with NP from IFV-A3-infected cells. And immunohistochemistry assays suggested that anti-NP1, anti-NP2, anti-NP3 antisera positively reacted with IFV-A3 or IFV-A1-infected MDCK cells, with titers of 1:640 to 1:1280.

CONCLUSIONThe recombinant NP of IFV-A3 would induce polyclonal antibodies with high titers in mice. The polyclonal antibodies would cross-react with IFV-A3 and IFV-A1. It is feasible to predict the antigenicity with systematical bioinformatics analyses and then induce anti-IFV antibodies with high dilutions, and it is possible to be utilized in the early detection and subtyping analyses of IFV-infections.

Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Influenza A virus ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology

OBJECTIVEHuman mannose-binding lectin (MBL) is a C-type serum lectin synthesized by the liver as an acute-phase protein. MBL can bind to glycoproteins terminated with mannose and N-acetylglucosamine present in the cell walls on a variety of microorganisms. Therefore, MBL appears to play an important role in the immune system. Low levels of MBL in human have been associated with a susceptibility to recurrent infections. MBL deficiency and low serum MBL levels are strongly associated with the presence of three point mutations at codon 52, 54 and 57 of exon 1 in the human MBL gene, and in Chinese population, the codon-54 mutation occurs at a frequency of 0.11 - 0.17. The data suggested that MBL insufficiency might also predispose to the development of autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). The possibility that Kawasaki disease (KD) is an infectious disease has been discussed and investigated for decades, in light of the implication that infections are involved in the pathogenesis of KD. It has been suggested that MBL insufficiency might predispose to the occurrence of KD. This study was aimed to investigate the genetic association of MBL codon-54 polymorphism in patients with KD, and to investigate possible associations with clinical manifestations of the disease.

METHODSThere were 95 patients with KD and 160 healthy subjects in the study. The genotype of MBL gene 54 codon was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Clinical characteristics and biochemical examination were also performed.

RESULTSThe genotype frequency of heterozygote (GGC/GAC) was significantly higher in KD group than that in healthy subjects (45.2% vs 25.0%, P < 0.01), and the allele frequency of GAC mutation was also higher in KD patients than that in control group (0.258 vs 0.138, P < 0.01). The variant allele (GAC) was markedly associated with KD (OR = 2.18, 95% CI = 1.38 approximately 3.44, P < 0.05). But there was no significant difference in the allele frequency of GAC between patients with and without coronary artery lesion (CAL) in KD cases (0.281 vs 0.246, P > 0.05). In addition, in cases of KD, more patients carrying the variant allele (GAC) had episodes of upper respiratory or gastrointestinal infections prior to the onset of KD than wild homozygotes (P < 0.01).

CONCLUSIONThe codon 54 polymorphism of MBL gene was associated with KD. It is possible that MBL gene codon 54 mutation might be related to the pathogenesis of KD.

Child ; Child, Preschool ; Codon ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Humans ; Infant ; Male ; Mannose-Binding Lectin ; genetics ; Mucocutaneous Lymph Node Syndrome ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Polymorphism, Restriction Fragment Length

OBJECTIVETo evaluate the immuno-effects of hepatitis B (HB) vaccination in adults.

METHODSFive groups were sampled by means of cluster sampling, and serum HBsAg, anti-HBs and anti-HBc were tested in every group at people aged from 18 to 50. Recombinant HB vaccine was injected to the ones that HBsAg, anti-HBs and anti-HBc were all negative. Concentration of anti-HBs in serum was tested after one year and three years of vaccination. Immuno-effects of recombinant HB vaccination in adults at different ages and between sexes, were then calculated.

RESULTSGood immuno-effects of recombinant HB vaccination in adults were noticed. After one year and three years of vaccination with 5 micro g recombinant HB vaccine, the anti-HBs positive rates were 82.76%, 70.77% while the serum concentrations of anti-HBs were 55.91 mIU/ml and 35.41 mIU/ml respectively. When 10 micro g was used, the concentrations were 83.74%, 72.22%, 56.89 mIU/ml and 30.29 mIU/ml respectively. The effects did not show significant differences between different doses on 10 micro g and of 5 micro g. Concentration of anti-HBs reduced when time went by. The factors such as age and sex influenced the effects of immunity on recombinant HB vaccination.

CONCLUSIONGood immunity could be obtained when recombinant hepatitis B was vaccinated in vulnerable population aged 18 to 50.

Adolescent ; Adult ; Female ; Hepatitis B ; prevention & control ; Hepatitis B Antibodies ; blood ; Hepatitis B Core Antigens ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; immunology ; Humans ; Male ; Middle Aged ; Vaccination ; Vaccines, Synthetic ; immunology

OBJECTIVECleidocranial dysplasia (CCD) is a dominantly inherited skeletal dysplasia caused by mutations in the osteoblast-specific transcription factor-encoding gene, core binding factor α1 (CBFA1). Over 90 mutations in CBFA1 gene have been published to date in 500 independent cases of CCD, including missense mutations, deletions, insertions, frameshift, and splice mutations. However, mutational screening of the CBFA1 gene is still far from saturation, and more novel mutations will be identified to enrich the insights into the molecular basis for the pathogenesis of CCD. The aim of this study was to explore the clinical and image features and detect the mutations of CBFA1 gene in two CCD families.

METHODIn this study, the clinical features were investigated in two CCD families, radiological and CT examinations regarding osseous malformation were carried out over the entire body of these patients with CCD. Blood (2 ml) was drawn from all affected individuals, unaffected family members and one hundred unrelated normal controls, Genomic DNA was extracted from whole blood with PureGene DNA extraction kit and PCR was performed with eight pairs of PCR primers for exons 0 to 7 of the CBFA1 gene. The mutations of CBFA1 gene were screened in these two CCD families.

RESULT(1) The clinical features of patients with CCD include delayed closure of fontanelles, frontal bossing, dysplasia of clavicles, late tooth eruption, and other skeletal anomalies. X-ray and CT examination showed the bulging calvarium, patent fontanelles, wide cranial sutures, multiple Wormian bones, dental dysplasia or aplasia of clavicles. (2) Two mutations were identified, one is novel missense mutation (c.1259C > T[p.T420I]) in CBFA1 gene exon 7, other (c.577C > T[p.R193X]) was reported in Chinese cases with CCD for the first time.

CONCLUSION(1) The clinical and image features of patients in two CCD families include delayed closure of fontanelles, frontal bossing, dysplasia of clavicles, late tooth eruption, and other skeletal anomalies. (2) The T420I and R193X mutations of CBFA1 were reported, expanding the spectrum of CBFA1 mutations causing CCD.

Child ; Child, Preschool ; Cleidocranial Dysplasia ; genetics ; pathology ; Core Binding Factor Alpha 1 Subunit ; genetics ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

OBJECTIVETo study the protective effects of lycopene (LP) on cerebral ischemia-reperfusion injury induced by focal cerebral ischemia and oxidative stress in rats.

METHODS48 male Sprague-Dawley (SD) rats were randomly assigned into five groups: A (20 mg/kg LP), B (5 mg/kg LP), C (salad oil), D (salad oil) and E (basic feed control). A, B and C groups were given LP or salad oil orally for 15 d, then cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion (MCAO) and D group was used as fake surgery control. The contents of reactive oxygen species (ROS), nitric oxide (NO), lactic acid (LD) and the activities of nitric oxide synthetase (NOS) in cortex were measured at 24 h after reperfusion. The levels of HIF-1alpha mRNA and Bcl-2 mRNA in hippocampi were determined by using reverse transcription polymerase chain reaction (RT- PCR) technique.

RESULTSROS levels of A, B, C, D and E groups were (114.23 +/- 18.91), (135.89 +/- 14.17), (171.37 +/- 25.76), (94.24 +/- 2.23) and (92.06 +/- 5.59) fluorescence intensity value/g protein, respectively (F = 9.038, P < 0.01); levels of NO were (6.60 +/- 0.77), (7.13 +/- 0.47), (8.38 +/- 0.80), (5.52 +/- 0.16) and (5.23 +/- 0.51) micromol/g protein respectively (F = 10.197, P < 0.01); levels of NOS were (0.817 +/- 0.016), (0.875 +/- 0.095), (1.030 +/- 0.101), (0.557 +/- 0.094) and (0.595 +/- 0.066) U/mg protein respectively (F = 14.555, P < 0.01); levels of LD were (0.381 +/- 0.069), (0.446 +/- 0.012), (0.576 +/- 0.059), (0.359 +/- 0.021) and (0.310 +/- 0.036) mmol/g protein respectively (F = 10.043, P < 0.01); HIF-1alpha mRNA expression levels in hippocampi were 0.865 +/- 0.274, 0.635 +/- 0.069, 0.491 +/- 0.067, 0.375 +/- 0.052 and 0.361 +/- 0.087, respectively (F = 40.520, P < 0.01); and Bcl-2 mRNA expression levels in hippocampi were 0.263 +/- 0.033, 0.330 +/- 0.028, 0.198 +/- 0.034, 0.304 +/- 0.039 and 0.236 +/- 0.025, respectively (F = 11.003, P < 0.01).

CONCLUSIONThe protective effects of LP may be related with its abilities of decreasing ROS and LD cumulation, alleviating inflammation and up-regulating the expression of protective genes.

Animals ; Antioxidants ; pharmacology ; Brain Ischemia ; metabolism ; Carotenoids ; pharmacology ; Hippocampus ; metabolism ; Infarction, Middle Cerebral Artery ; metabolism ; Lactic Acid ; metabolism ; Male ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Reperfusion Injury ; metabolism

OBJECTIVEKawasaki disease (KD) is an acute febrile vasculitic syndrome of unknown etiology that preferentially affects coronary artery. It has been suggested that proinflammatory cytokines like tumor necrosis factor alpha (TNF-alpha) and interleukin-10 (IL-10) are key players during acute KD. Recently, the polymorphisms relative to major transcriptional start site of TNF-alpha and IL-10 gene were shown to influence the level of TNF-alpha and IL-10 production in vitro. This study was aimed to investigate the genetic association of TNF-alpha and IL-10 promoter polymorphisms in juvenile patients of Han nationality with KD, and to investigate the possible associations with clinical manifestations of the disease.

METHODSFour polymorphism sites of TNF-alpha and IL-10 gene promoter regions from 96 children with KD were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). One hundred and sixty age-matched normal children of the Han nationality were used as control. All patients accepted Doppler echocardiography examination in order to differentiate coronary artery lesions.

RESULTSThere was significant difference in allele frequencies of -308 (A/G) site of the TNF-alpha gene between children of the Han nationality and those of Japanese and Caucasian in America. There were significant differences in the allele frequencies of -1082 (G/A), -819 (C/T) and -592 (A/C) of IL-10 gene between children of the Han nationality and their British Counterparts (P < 0.01). There was no significant difference in allele frequencies of -308 (A/G) site of TNF-alpha gene between children with KD and normal controls. There was no significant difference in the haplotypes and the allele frequencies of the above three sites of IL-10 between the two groups. However, when clinical features were examined, the genotype frequency of TNF-alpha-308A was significantly higher in IVIG-resistant KD patients than that of TNF-alpha-308G genotype (67% vs 5%, chi(c)(2) = 90.48, P < 0.01). The genotype of TNF-alpha-308A was closely associated with IVIG-resistant KD (P < 0.01, relative risk 42.25, 95% confidence interval 15.81-112.88). The haplotype frequency of IL-10 -1082A/-819T/-592A was also higher in patients with coronary artery lesion (CAL) caused by KD than those of Non-ATA haplotype (52% vs 20%, chi(2) = 18.36, P < 0.01). The haplotypes of IL-10 -1082A/-819T/-592A was significantly associated with CAL caused by KD (P < 0.01, relative risk 4.26, 95% confidence interval 2.20-8.25).

CONCLUSIONThe genotype of TNF-alpha-308A is one of the important factors that probably influence the therapeutic effect of KD. The haplotypes (-1082/-819/-592) of IL-10 gene promoter might be related to the pathogenesis of coronary artery complication of KD and -1082A/-819T/-592A haplotypes might be regarded as a genetic marker of risk factor for coronary artery lesion in KD.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Interleukin-10 ; genetics ; Male ; Mucocutaneous Lymph Node Syndrome ; genetics ; pathology ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic ; genetics ; Tumor Necrosis Factor-alpha ; genetics