Objective To evaluate the contribution of gyrA and parE detection in Ureaplasma genotyping.Methods Sixty Ureaplasma isolates were selected with the Mycoplasma IST kit.The gyrA and parE were amplified by PCR.The DNA was sequenced and compared with the corresponding sequences in GenBank.Results The nucleotide sequence of gyrA had 100% identity in serovar 1,3,6,14 and 100%identity in serovar 2,4,5,7～13,too.But the sequence had 91%identity between the two groups.The nucleotide sequence of parE had 98%～99% identity in serovar 1,3,6,14.And it had 100% identity in erovar 2,5,7,8,11 and 100% identity in serovar4,12,13.But it had only 90% identity between the two groups.Ureaplasma parvum(Up),Ureaplasma urealyticum(Uu)and Up+Uu infection were found 68.3%(41/60),21.7%(13/60)and 10%(6/60) of clinical specimens,respectively.In Up isolates,serovar 3 was 48.8%(20/41).Conclusion Ureaplasma can be divided into two genotypes(Up and Uu)by gyrA analysis.And Up can be divided into four subtypes which correspond to serovar 1,3,6,14,respectively.Serovar 3 is the main isolate in our research.

OBJECTIVETo investigate the chemical constituents of dried whole plants of Artemisia annua.

METHODThe chemical constituents were isolated by repeated silica gel chromatography, medium pressure column chromatography, and semi-preparative HPLC, and their structures were elucidated by spectroscopic analyses and comparison of NMR data with those reported in literature.

RESULT15 compounds were isolated and identified to be 5-O-[(E)-Caffeoyl] quinic acid(l), 1,3-di-O-caffeoylquinic acid(2), 4 5-di-O-caffeoylquinic acid(3), 3, 5-di-O-caffeoylquinic acid (4), 3, 4-di-O-caffeoylquinic acid (5), methyl-3,4-di-O-caffeoylquinic acid(6), methyl-3,5-di-O-caffeoylquinic acid(7), 3,6'-O-diferuloylsucrose(8), 5'-β-D-glucopyranosyloxyjasmonic acid(9), Scopoletin(10), scoparone (11), 4-O-β-D-glucopyranosyl-2-hydroxyl-6-methoxyacetophenone (12), chrysosplenol D (13), casticin (14), chrysosplenetin(15).

CONCLUSIONCompounds 2, 6, 8 and 9 are obtained from the Artemisia genus for the first time. Compounds 7 and 15 are obtained from this plant for the first time.

Artemisia annua ; chemistry ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Medicine, Chinese Traditional ; Plants, Medicinal ; Quinic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Silica Gel

1,2,3,4,6-penta-O-galloyl-D-glucose (PGG) is one of the main active compounds of Guizhi Fuling capsule. Molecularly imprinted polymers (MIP) have high affinity toward template molecules synthesized by molecularly imprinted technology for its specific combined sites, which can overcome the shortcoming of traditional separation methods, such as complex operation, low efficiency, using large quantity of solvent and environmental pollution. In this paper, surface molecularly imprinted polymer (SMIP) was prepared by surface imprinting with PGG as the template molecule. Its adsorption capacity was measured by the scatchard equation. The separation of PGG from Guizhi Fuling capsule at preparatived scale was achieved with molecularly imprinted polymer as stationary phase and the purity was 90.2% by HPLC. This method can be used to prepare PGG from Guizhi Fuling capsule with large capacity and is easy to operate. It provides a new method for efficient separation and purification for other natural products.
Adsorption ; Capsules ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Hydrolyzable Tannins ; chemistry ; isolation & purification ; Molecular Imprinting ; Polymers ; chemical synthesis ; chemistry

OBJECTIVETo study whether N, N'-di-(m-methylphenyi)-3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboamide (ZGDHu-1) inhibits proliferation and induces apoptosis in human lung carcinoma cell line EBC-1 cells and its molecular mechanism.

METHODSDifferent concentrations of ZGDHu-1 and different times of culture were used to treat EBC-1 cells in vitro. The inhibition of proliferation was measured by BrdU-ELISA. Cell apoptosis was detected by Annexin V/PI staining and cellular DNA fragmentation ELISA. Phosphorylated p38MAPK and STAT3 were examined by flow cytometry. The protein expressions of bcl-2, bax, p53, Fas, and caspase-3 were detected by Western blot analysis.

RESULTSZGDHu-1 inhibited EBC-1 cell proliferation within a certain range of treating times and does, with a 24 h IC(50) of (295 ± 25) ng/ml, 48 h of (112 ± 8) ng/ml and 72 h of (23 ± 2) ng/ml. The EBC-1 cell apoptosis was confirmed by Annexin V/PI labeling and cellular DNA fragmentation ELISA in a dose-related manner. When EBC-1 cells were treated with 50, 200, and 500 ng/ml ZGDHu-1 for 48 h, the expression rates of phosphor-p38MAPK protein were 67.4%, 88.2%, 91.1%, respectively, and that of the control was 10.6%. That of STAT3 protein were 56.5%, 43.6% and 34.6%, respectively, and that of the control was 89.1%. The expression of bax, p53 and Fas protein was significantly increased, that of bcl-2 was not changed, and that of caspase-3 was significantly decreased by the ZGDHu-1 treatment.

CONCLUSIONZGDHu-1 can inhibit proliferation and induce apoptosis in EBC-1 cells. The mitochondrial pathway mediated by Fas may be one of its mechanisms. The apoptosis of EBC-1 cells may associate with up-regulation of phosphor-p38MAPK and down-regulation of phosphor-STAT3 in the cells.

Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Fragmentation ; Heterocyclic Compounds, 1-Ring ; administration & dosage ; pharmacology ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; STAT3 Transcription Factor ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo study the anti-complementary phenolic acids from Lonicera japonica.

METHODThe anti-complementary activity-directed isolation was carried out with the hemolysis test as guide. All isolation was evaluated for their in vitro anti-complementary activities. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR, 13C-NMR data.

RESULTFourteen compounds were isolated from the EtOAc fraction of L. japonica extracts, including 8 phenolic acids: 5-O-caffeoylquinic acid (1), chlorogenic (2), 4-O-caffeoylquinic acid (3), 3,5-di-O-caffeoylquinic acid (4), 4,5-di-O-caffeoylquinic acid (5), 3,4-di-O-caffeoylquinic acid (6), caffeic acid (7) and methyl caffeate acid (8); 3 iridoids: secologanoside (9), sweroside (10) and secoxyloganin (11); and 3 flavonoids: luteolin (12), quercetin (13) and kaempferol (14). Compounds 1-9 and 11-14 showed anti-complementary activity in different extents and 3,5-di-O-caffeoylquinic acid (4) exhibited the most significant activity against the classical pathway.

CONCLUSIONCompound 14 is obtained from this plant for the first time, phenolic acids are the main anti-complementary constituents of L. japonica and 3,5-di-O-caffeoylquinic acid(4) is a potential complement inhibitor with strong activity, which worthy to be studied further in the future.

Complement Inactivating Agents ; chemistry ; isolation & purification ; pharmacology ; Hydroxybenzoates ; chemistry ; isolation & purification ; pharmacology ; Lonicera ; chemistry

The present study was aimed to analyze the immunogenicity of recombinant human zona pellucida-3 peptides (r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348)) expressed in E. coli through immunizing rabbits, and to evaluate the efficacy of their polyclonal antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding respectively. Male New Zealand rabbits were immunized using r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) as antigen respectively, which was purified through an improved method of preparative gel polyacryulamide gel electrophoresis. The antibody response level of r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) immunogen in rabbits was determined by ELISA using mouse ZP3-5 (amino acid sequence(137 approximately 150) being completely conserved with huZP3(138 approximately 151) sequence) and specific huZP3-14 (amino acid sequence(327 approximately 340)) synthetic peptides as coating antigens respectively. The immunoreactivity and specificity of the anti-r-huZP3a(22 approximately 176) and anti-r-huZP3b(177 approximately 348) antisera with each r-huZP3 peptides, were tested by immunoblot and immunohistochemistry (using native huZP and human ovary section) respectively. A competitive hemizona assay (HZA) was used to evaluate the efficacy of the antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding. Both r-huZP3 peptides were able to induce higher antibody titers in rabbits. Each antiserum could specifically recognize or bind to each target r-huZP3 peptide expressed in E. coli and native human ZP in vitro. The antisera also inhibited sperm-egg binding in the HZA. These results show that r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) are of strong immunogenicity. They can be used to develop a kit for detecting whether there are autoantibodies to zona pellucida in unexplained infertile women, and their antisera might be useful tools for determining minimal B-cell epitope sequences of several known huZP3 epitope peptides.
Animals ; Egg Proteins ; biosynthesis ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Immune Sera ; immunology ; Immunization ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; immunology ; Rabbits ; Receptors, Cell Surface ; biosynthesis ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Sperm-Ovum Interactions ; immunology ; Zona Pellucida Glycoproteins

Objective To analyze the status of health literacy and its relationship with chronic diseases and self-reported health among residents in Qingdao, discuss the influence of health literacy on health status and provide the scientific basis for the development of health education strategies and measures. Methods The stratified multistage and probability proportionate to population size sampling(PPS sampling) method was adopted in this investigation. In 2017, a questionnaire survey was conducted among 16 700 permanent residents aged 15-69 from 10 districts in Qingdao. Results The overall level of health literacy status was 15.92%, the prevalence rate of chronic diseases was 19.31%, the self-reported health proportion of good, fair and poor were 81.68%, 12.12% and 1.71% among residents in Qingdao. Logistic regression analysis showed that, after adjusting for urban and rural areas, gender, age, education, income and occupation, health literacy was a protective factor for people with chronic diseases and self-evaluated health(OR=1.232，P=0.003;OR=1.159，P=0.033). Three aspects of health literacy were correlated with chronic diseases and self-reported health among people (all P<0.05). Conclusions Health literacy is positively correlated with the health status of residents. The improvement of health literacy is an important way to enhance the health status of residents.

A 558 bp cDNA fragment was amplified by RT-PCR from adult Schistosoma japonicum(Chinese strain) mRNA with a pair of primers that were designed according to published Sj21.7p gene encoding 21.7 kD protein of Schistosoma japonicum(Philippines strain). Sequence analysis indicated that this frame, named Sj21.7 (Ch), with 99% homology to Sj21.7 p, contained a complete open reading fragment (ORF) of 21.7 kD protein gene of Schistosoma japonicum(Chinese strain). The amino acid sequence shared 98% homology with 21.7 kD protein of Schistosoma japonicum. This fragment was cloned into the expression vector pET28a (+) and subsequently expressed in Escherichia coli with IPTG induction. SDS-PAGE analysis revealed that the molecular weight of this expressed product was 25.4 kD. Western blotting showed that the recombinant protein reacted well with the rabbit serum immunized with Sj worm antigen, indicating that this expressed product had good antigenicity.
Amino Acid Sequence ; Animals ; Cloning, Molecular ; Helminth Proteins ; biosynthesis ; chemistry ; genetics ; Molecular Sequence Data ; Molecular Weight ; Open Reading Frames ; Rabbits ; Recombinant Proteins ; biosynthesis ; chemistry ; immunology ; Schistosoma japonicum ; genetics ; Sequence Homology

OBJECTIVETo estimate the prevalence of parvovirus B19 infection in Chinese Xiamen area blood donors.

METHODSBlood samples from blood donors were tested for detection of parvovirus B19 DNA and antibody. The direct sequencing and genetype analysis of B19 DNA positive samples were performed.

RESULTSSix out of 10452 samples were B19 DNA positive. The viral loads of the 6 samples were between 3.59×10-1.07×10IU/ml; the positive rate of B19-IgM was 4.64%(50/1078) and B19-IgG was 16.79%(181/1078). The positive rate of B19-IgG increased with ages, and was not related with the sex.

CONCLUSIONThe overall prevalence of parvovirus B19 infection in blood donors is lower in Chinese Xiamen area than that in other areas, however, there is still a certain percentage of viremia in donors and the attention should be paid to blood safety in the future work.

Acute Disease ; Adult ; Cerebral Infarction ; diagnosis ; diagnostic imaging ; Fabry Disease ; diagnosis ; diagnostic imaging ; Humans ; Magnetic Resonance Imaging ; Male