There are reports about the chemical compounds of Ciwujia herbs, but with no study report about the chemical material basis of Ciwujia injection (CWJI). In this study, LC-MS(n) and LC-Q-TOF-MS techniques were adopted for a qualitative analysis on phenylpropanoids in CWJI. The Ultmate XB-C18 column (4.6 mm x 250 mm, 5 microm) was adopted and eluted with the mobile phase of 0.5% formic acid-water and acetonitrile, with the flow rate at 0.8 mL x min(-1) and the column temperature at 20 degrees C. Based on the data of high-resolution and multi-stage MS, control products and literatures, altogether 54 phenylpropanoids were identified in Ciwujia Injection, including 34 phenylpropanoids, 16 ligans and 4 coumarins. Among them, 28 were reported for the first time in Ciwujia, and 14 compound structures were identified in comparison with the control products. The method established in this study could be used to simply and rapidly identify phenylpropanoids in CWJI. The findings provide scientific data for defining the chemical material basis of CWJI.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Mass Spectrometry ; methods ; Molecular Structure

OBJECTIVETo estimate the feasibility of 16-multidetector spiral computed tomography (16-MDCT) on detecting coronary plaques in comparison with intravascular ultrasound (IVUS).

METHODSSixty-eight patients suspected of coronary heart diseases were examined by 16-MDCT, quantitative coronary angiography (QCA) and IVUS. Coronary stenosis was defined as lumen diameter reduction (DS) >or= 50%. Hounsfield units (HU) were used to determine different types of plaques: soft plaque (or= 120 HU).

RESULTSCompared to QCA, the sensitivity and the specificity for patients with DS >or= 50% were 91.8% (112/122) and 97.8% (556/568) respectively, the positive and negative predictive value were 90.3% (112/124) and 98.2% (556/566) respectively. In 96 plaques evaluated both by 16-MDCT and IVUS, 20 and 21 soft plaques, 37 and 36 fibrous plaques, 39 and 38 calcified plaques were identified by 16-MDCT and IVUS respectively. HU value of soft (11 +/- 36), fibrous (83 +/- 20), and calcified (292 +/- 80) plaques were significantly different (P < 0.05).

CONCLUSIONSNoninvasive 16-MDCT could correctly estimate coronary stenosis and coronary plaques compositions.

Aged ; Atherosclerosis ; diagnostic imaging ; Coronary Angiography ; Female ; Humans ; Male ; Middle Aged ; Tomography, Spiral Computed ; methods ; Ultrasonography, Interventional

OBJECTIVETo study whether anxiety and depression are associated with the development of neurogenic urination in children.

METHODSA total of 136 9 to 12-year-old children with neurogenic urination (case group) and 136 age-matched healthy children (control group) were enrolled. The Screen for Children Anxiety Related Emotion Disorders (SCARED) and Depression Self-rating Scale for Children (DSRSC) were used to evaluate the psychological status. The incidences of anxiety and depression as well as the SCARED and DSRSC scores were compared between two groups. Logistic regression analysis model was used to evaluate the relationship between psychological status and the development of neurogenic urination.

RESULTSThe case group was found to have a higher incidence of anxiety and depression compared with the control group (P<0.01). The SCARED score in the case group (28.1 ± 8.6) increased significantly compared with 14.4 ± 4.9 in the control group (P<0.01). The DSRSC score in the case group was also significantly higher than in the control group (13.5 ± 4.8 vs 9.1 ± 3.2; P<0.01). The logistic regression analysis showed that the children with anxiety (SCARED-score>23) had a 1.224-fold increased risk for the development of neurogenic urination compared with the children with the SCARED-score≤23 and that the children with depression (DSRSC-score&ge;15) had a 1.148-fold increased risk for the development of this disorder.

CONCLUSIONSAnxiety and depression participate in the development of neurogenic urination in school children.

Anxiety ; complications ; Child ; Depression ; complications ; Female ; Humans ; Logistic Models ; Male ; Urination Disorders ; etiology ; psychology

OBJECTIVETo investigate the detection limit of multicolor combinational probe coding real-time PCR (MCPC-PCR) in detection of Salmonella and Staphylococcus aureus suspended in the food samples, and to apply MCPC-PCR to detect the samples of food poisoning.

METHODSSeries concentration of bacterium suspension (10(1) - 10(9) CFU/ml) was prepared by using 22 simulated samples including fresh meat and cakes and then MCPC-PCR was applied to detect Salmonella and Staphylococcus aureus in 22 samples. Enrichment broth of 101 frozen samples and 5 early patients' anal swabs in food poisoning cases were detected after the DNA samples were extracted.

RESULTSThe limits of MCPC-PCR assay in detecting Salmonella and Staphylococcus aureus were about 10(2) copies/test; 101 frozen enrichment broth of samples in food poisoning cases were detected by MCPC-PCR assay, of 23 positive samples, 18 were confirmed by bacteriology techniques; 96 samples detected by MCPC-PCR and bacteriology techniques had the same results, and the coincidence rate was 95.05%. Anal swabs, collected from 5 of early patients in a food poisoning case gave a clue to be Vibrio parahaemolyticus by MCPC-PCR assay and then were perfectly consistent with bacteriology assay.

CONCLUSIONAs a method of high sensitivity and good specificity, MCPC-PCR assay can quickly and conveniently detect multiple pathogens existing in food samples, therefore we recommend it to be used in rapidly screening or simultaneous detection of food-borne diseases.

Bacteriological Techniques ; methods ; Food Contamination ; analysis ; Food Microbiology ; Molecular Probe Techniques ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Salmonella ; genetics ; isolation & purification ; Sensitivity and Specificity ; Staphylococcus aureus ; genetics ; isolation & purification

The secondary structure of Capsid protein was predicted by the methods of Chou-Fasman,Garnier-Robson and Karplus-Schultz based on the sepuence of capsid protein gene of Swine Vesicular Disease Virus (SVDV) and hydrophilicity. Surface probility plot and antigenic index for capsid protein were obtained by the methods of Kyte-Doolittle, Emini and Jameson-wolf, respectively, Combining the results according to these methods, the B cell epitopes for capsid protein of SVDV were predicted. The results showed that there are much flexible region such as coil region and turn region in capsid protein of SVDV, there are more predominant B cell epitopes in VP1 than in VP2 and VP3. This study would be helpful for identification of B cell epitopes for capsid protein using experimental methods and research of reverse vaccine of SVDV.

OBJECTIVETo prepare the oral solution of egg yolk hepatitis B virus (HBV)-specific transfer factor (EYHBV-TF) and evaluate its immunological activity as an immune regulator against hepatitis B.

METHODSFrom hens immunized with the Hepatitis B vaccine the egg yolk was isolated to extract the specific transfer factor EYHBV-TF, and its physicochemical properties were examined. Leukocyte adhesion inhibition test (LAI) was performed to detect the immunogenic activity of EYHBV-TF. The solution of EYHBV-TF was then administered orally in normal mice, and the specific cellular immune activity induced was assayed with delayed type skin hypersensitivity test (DTH), with the non-specific immune activity assessed with immune organ index. The immune responses induced by oral EYHBV-STF solution were compared with those by EYHBV-STF injection and by different dosages (injection and oral) of porcine spleen HBV-specific transfer factor (PSHBV-STF), porcine spleen nonspecific transfer factor, and egg yolk extracts from non-immunized hens.

RESULTSThe prepared EYHBV-STF oral solution, which met the standards for biological products, could inhibit leukocyte adhesion in vitro and significantly enhance mouse foot pad swelling, demonstrating its capability of transferring antigen-specific delayed type hypersensitivity reactions to naive recipient. EYHBV-STF oral solution also significantly improved the immune organ index in mice (P<0 01) with similar effects to those caused by EYHBV-STF injections and by PSHBV-STF injection and oral solution.

CONCLUSIONOrally administered EYHBV-STF and EYHBV-STF injection both possess hepatitis B antigen-specific cellular immune activity and can significantly enhance specific cellular immune responses.

Animals ; Chickens ; Egg Yolk ; chemistry ; Hepatitis B ; drug therapy ; Hepatitis B Antigens ; Hepatitis B virus ; drug effects ; Immunity, Cellular ; Immunization ; Mice ; Swine ; Transfer Factor ; administration & dosage ; pharmacology

Objective To compare the effects of three different methods for extracting short RNA-DNA hybrids, including the TRI reagent method , the phenol saturated with water method and the phenol saturated with Tris buffer method in order to facilitate studies on the biological function of RNA-DNA hybrids .Methods Short RNA fragments modifiedwith FAM at the 5′end and those modified with Cy 5 at the 5′end were synthesized .RNA and DNA fragments were annealed to form RNA-DNA hybrids.They were extracted with the above-mentioned 3 methods respectively .The extracted products were analyzed with electrophoresis .Results and Conclusion Short RNA-DNA hybrids can be extracted by the phenol saturated with water method and by the phenol saturated with Tris buffer method .The results can help study the function of short RNA-DNA hybrids .

OBJECTIVETo evaluate the clinical performance of a probe melting analysis (PMA)-based real-time PCR detection kit in rapid detection of rifampin-resistant mutations in Mycobacterium tuberculosis (MTB).

METHODSThe specificity of the assay was evaluated by detecting 37 non-tuberculous mycobacteria (NTM), and the detection limit of the method was evaluated by genomic DNA of a standard strain H37Rv. Finally, 962 clinical isolates were analyzed with the PMA assay by detecting mutations in rifampin resistance-determining region (RRDR) of rpoB gene, and results were verified with DNA sequencing.

RESULTSAmong 37 NTM strains, three strains showed drug resistant mutation signals. The PMA method could detect down to 30 bacteria per reaction. Sample analysis showed that 186 of 962 isolates were mutants, 751 isolates were wild type and 25 isolates failed to give amplification signals. Among the mutant samples detected, 112 samples from November 2009 to April 2010 were further analyzed by sequencing, as well as 200 wild-type samples. The results showed a complete agreement with the PMA assay except for 5 samples failed in sequence analysis.

CONCLUSIONThe PMA assay is rapid, accurate and easy-to-use, and thus can be used for detection of rifampin-resistant in clinical isolate samples.

Bacterial Proteins ; genetics ; Base Sequence ; DNA Mutational Analysis ; DNA, Bacterial ; genetics ; DNA-Directed RNA Polymerases ; Genotype ; Limit of Detection ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Mutation ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

OBJECTIVETo observe the intervention effect of Leihong Granule (LG) in in-stent restenosis (ISR) after endovascular therapy for lower extremity arterial occlusive diseases (LEAOD).

METHODSRecruited 80 LEAOD patients who successfully underwent endovascular therapy (balloon dilation and stent implantation) were randomly assigned to two groups, the control group and the LG group, 40 in each group. Patients in the control group received basic treatment, while those in the LG group additionally took LG for 3 months. Plasma levels of IL-10, IL-18, CRP, and the intima-media thickness (IMT) of lower extremity artery were observed in the two groups between and after treatment. The rate of stent patency, ABI, intermittent claudication, rest pain, and the incidence of amputation the two groups were recorded and observed in the two groups.

RESULTSIn the control group, serum levels of IL-10, IL-18, CRP, and IMT were significantly higher one month after surgery than before surgery (P < 0.05). There was no significant difference in serum levels of IL-10, IL-18, CRP, or IMT between the two groups before surgery (P > 0.05). These indices were obviously lower in the LG group than in the control group after surgery (P < 0.05). Compared with the control group, the incidence rates of intermittent claudication and the rest pain at 6 months and 12 months after surgery significantly decreased (P < 0.05). The stent patency rate at 6 months and 12 months after surgery, and ABI were significantly higher than those of the control group (P < 0.05). There was no statistical difference in the amputation rate between the two groups (P > 0.05).

CONCLUSIONLG might effectively improve ischemic symptoms of affected limbs possibly through lowering the ISR rate after endovascular therapy for LEAOD through preventing immunosuppressive actions.

Aged ; Aged, 80 and over ; Arterial Occlusive Diseases ; therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Graft Occlusion, Vascular ; therapy ; Humans ; Interleukin-10 ; blood ; Interleukin-18 ; blood ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Phytotherapy ; Stents ; Treatment Outcome

Receptors play a crucial role in determining the host specificity and tissue tropism of virus. Foot-and-mouth disease virus(FMDV)has been showed to use four integrins, ?v?1, ?v?3, ?v?6 and ?v?8 as receptors to initiate infection and ?v?6 functions as the major receptor.The cDNA encoding bactrian camel integrin ?6 from the lung tissue was cloned and sequenced. The 2367bp cDNA of bactrian camel integrin ?6 encodes a polypeptide of 788 amino acids consisting of a 26-residue putative signal peptide, a 681-residue ectodomain with 8 potential N-linked glycosylation sites and 58 cysteine residues, a 29-residue transmembrane domain, and a 52-residue cytoplasmic domain with a NPLY motif and 1 potential N-linked glycosylation site. The nucleotide sequence similarity of integrin ?6 between bactrian camel and cattle, pig, sheep, human, mouse, Norway rat is 91.1%、91.8%、90.6%、90.5%、83.7%、84.1%, and the amino acid sequence similarity is 94.3%、93.4%、93.4%、93.7%、88.7%、88.6%, respectively. The bactrian camel ?6 gene exhibited the higher sequence homology with the ?6 gene of cattle, pig and sheep, indicating their close genetic relationships. It is possible that host tropism of FMDV may related to divergence in ?6 receptors among different species.