Abstract: Objective To establish an animal model of BALB/c mice infected with Burkholderia pseudomallei through the nose (inhalation route), provides a reliable animal model for the follow-up studies on the virulence of melioidosis and the pathogenesis of acute melioidosis. Methods　The experiment was carried out through infecting with Burkholderia pseudomallei through the nose (inhalation route). The pathophysiological response, visceral pathological damage and bacterial colonization of the mice infected with Burkholderia pseudomallei were observed by gross anatomy, histopathology and tissue homogenate count, and the biological characteristics of the mouse model of acute melioidosis were analyzed accordingly. Then we compared the physiological responses in BALB/c mice between the Burkholderia pseudomallei and non-pathogenic Burkholderia thailandensis. Results　In the model of acute nasal infection with Burkholderia thailandensis, most death happened between the 3rd to 5th day after infection, 3×105-3×106 CFU was the suitable dose for acute fatal melioidosis model of BALB/c mice, and the medium lethal dose was about 3×104-3×105 CFU. Both gross anatomy and tissue HE staining showed that abscesses or necrosis were found in the lung, spleen and liver, especially in the spleen and lung, which was positively correlated with the challenge dose. Viable bacteria was isolated from the blood, lung, spleen and liver of Burkholderia pseudomallei-infected mice, and the bacteria account colonization was related to tissue specificity. The concentration of live bacteria isolated from in the blood was the highest [Log2 value: (10.28±0.34) CFU/mL], and the organ with the maximum quantity of bacteria was the lung [Log2 value: (7.54±2.11) CFU/total organ]. It has been reported that the biological effects of Burkholderia pseudomallei and its homologous non-pathogenic Burkholderia thailandensis were similar at the cellular level, like multi-nuclear giant cell formation and active intracellular replication, while it is still unclarrified in the differences of virulence in mice. In this study, it was proved that Burkholderia thailandensis was not fatal to mice even at a high dose (8×107CFU), or detected from mice infected with it via nasal. Conclusion　We successfully established a reliable BALB/c mouse model (acute lethal model) of melioidosis via nasal infection, described its biological characteristics, and identified the different biological responses between Burkholderia pseudomallei and its homologous non-pathogenic Burkholderia thailandensis in mice.

Blood Cell Count ; Bone Marrow Cells ; metabolism ; Child ; Cord Blood Stem Cell Transplantation ; Cytokines ; therapeutic use ; Histocompatibility Testing ; Humans ; Male ; Myelodysplastic Syndromes ; blood ; therapy ; Treatment Outcome

Extraction process trajectory of Danhong injection by near-infrared (NIR) spectroscopy in conjunction with multivariate data analysis techniques was developed for in-line monitoring of extraction process. To capture the variation of batch process, the use of score, Hotelling T2 and DModX control charts was investigated for real-time monitoring of extraction process. Various abnormal behaviors of the test batches were detected in time by comparing the extraction process trajectory. It was concluded that the process trajectory for in-line quality control based on NIR spectroscopy was a feasible technology tool of the total process quality control during traditional Chinese medicine (TCM) manufacturing process.
Chemistry, Pharmaceutical ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Online Systems ; Quality Control ; Spectroscopy, Near-Infrared ; methods

Ten compounds, including seven sesquiterpenes, two phenols and one phenylpropanoid, were isolated from the roots of Illicium majus by means of silica gel, ODS, Sephadex LH-20, and preparative HPLC. On analysis of MS and NMR spectroscopic data , their structures were established as cycloparviflorolide (1), cycloparvifloralone (2), tashironin (3), tashironin A (4), anislactone A(5), anislactone B (6), pseudomajucin (7), syringaldehyde (8), methyl-4-hydroxy-3, 5-dimethoxybenzoate (9), and (E)-3-methoxy-4,5-methylenedioxycinnamic alchol (10), respectively. Compounds 1-4 and 8-10 were first isolated from this plant. In the in vitro assays, at a concentration of 1.0 x 10(-5) mol x L(-1), compounds 5 and 6 were active against LPS induced NO production in microglia with a inhibition rate of 75.31% and 53.7%, respectively.
Drugs, Chinese Herbal ; analysis ; chemistry ; Illicium ; chemistry ; Organic Chemicals ; analysis ; chemistry ; Plant Roots ; chemistry

BACKGROUNDTo investigate the dynamics of amyloid fiber formation of yeast (Saccharomyces cerevisiae) prion protein Sup35NM under the native condition to provide materials and clues for the elucidation of amyloid fiber formation.

METHODSThe Sup35NM gene was cloned and expressed in E. coli. The recombinant Sup35NM protein was purified under denaturing conditions through Nickel-Sepharose chromatography. Aliquots were removed at designated time points for transmission electron microscopy (TEM), circular dichroism (CD) spectra, protease K resistance assay, as well as thioflavin T (ThT) binding assay.

RESULTSThe Sup35NM expressed and purified under denaturing conditions. The morphological alteration of the Sup35NM in PBS (pH7.4) during the protein aggregation and amyloid fiber formation was visualized by TEM. The CD assay showed that the course of amyloid fiber formation underwent a conformational shift from alpha-helix to beta-sheet. The fibers had higher capacity of resistance to protease K digestion compared to the monomers. ThT fluorescence assay displayed a rapid growth phase before reaching a final equilibrium phase during the fiber formation, and the higher concentration of Sup35NM could greatly accelerate the fiber formation in vitro.

CONCLUSIONYeast prion protein Sup35NM forms amyloid readily under native conditions in vitro. The dynamics of Sup35NM amyloid formation may provide supporting evidences for the nucleating polymerization models of amyloid fiber formation.

Amyloid beta-Peptides ; genetics ; metabolism ; ultrastructure ; Electrophoresis, Polyacrylamide Gel ; Endopeptidase K ; metabolism ; Kinetics ; Microscopy, Electron ; Peptide Termination Factors ; Prions ; genetics ; metabolism ; ultrastructure ; Protein Binding ; Recombinant Proteins ; metabolism ; ultrastructure ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; metabolism ; ultrastructure ; Thiazoles ; metabolism

Objective To explore the changes of serum levels of tumor necrosis factors (sTNF-α and sTNF-β) and their soluble receptors (sTNF-R1 and sTNF-R2), and analyze their relationship with sleep quality and memory in patients with chronic insomnia disorder (CID). Methods Forty-four CID patients and 39 normal controls were enrolled. Pittsburgh Sleep Quality Index (PSQI) and Nine-Box Maze Test were used to assess the insomnia severity and memory functions, respectively. The serum levels of sTNF-α, sTNF-β, sTNF-R1 and sTNF-R2 were examined using protein-chip technology. Results Compared to the controls, CID patients had significantly higher number of errors in spatial working (Z=5.362, P<0.001) and object recognition memories (Z=3.260, P=0.001) in the Nine-Box Maze. In addition, CIDpatients had higher levels of sTNF-αand sTNF-β (Ps<0.001), and lower levels of sTNF-R1 and sTNF-R2 (Ps<0.001). The Spearmen correlation analysis showed that the levels of sTNF-α and sTNF-β were positively correlated with the scores of PSQI (Ps<0.001), whereas the levels of sTNF-R1 and sTNF-R2 were negatively correlated with the scores of PSQI (Ps<0.001). In the CID patients, sTNF-α levels were positively correlated with the errors in both spatial working (γ= 0.380, P=0.017) and object recognition (γ= 0.349, P= 0.030) memories, whereas sTNF-β levels were only positively correlated with the error in spatial working memory (γ=0.414, P=0.009). The levels of sTNF-R1 and sTNF-R2 were not correlated with memory performance (Ps>0.05). Conclusion CID patients have increased levels of sTNF-αand sTNF- whereas have decreased levels of R1 and R2. The elevated sTNF-α and sTNF-β levels are correlated with memory disorders in CID patients.

Castleman's disease is a slowly progressive and rare lymphoproliferative disorder. Here, we report a 55-year-old woman with superior mediastinal Castleman's disease being misdiagnosed for a long term. We found a 4.3 cm mass localized in the superior mediastinum accompanied with severe clinical symptoms. The patient underwent an exploratory laparotomy, but the mass failed to be totally excised. Pathologic examination revealed a mediastinal mass of Castleman's disease. After radiotherapy of 30 Gy by 15 fractions, the patient no longer presented previous symptoms. At 3 months after radiotherapy of 60 Gy by 30 fractions, Computed tomography of the chest showed significantly smaller mass, indicating partial remission. Upon a 10-month follow-up, the patient was alive and free of symptoms.
Antigens, CD20 ; metabolism ; Castleman Disease ; diagnosis ; immunology ; pathology ; radiotherapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Mediastinal Diseases ; diagnosis ; immunology ; pathology ; radiotherapy ; surgery ; Mediastinum ; diagnostic imaging ; pathology ; Middle Aged ; Multimodal Imaging ; Positron-Emission Tomography ; Radiotherapy, Intensity-Modulated ; Tomography, X-Ray Computed

OBJECTIVETo determine the prevalence of herpes simplex virus (HSV) and its correlates among HIV/AIDS patients in a county of Shanxi.

METHODSAll HIV-infected patients in a county in Shanxi province who were receiving antiretroviral treatment (ART) were included in this study. Participants were interviewed using standard questionnaires. Serum samples were tested to determine HSV-1 and HSV-2 infections.

RESULTSA total of 195 AIDS patients were recruited and 195 blood samples were collected. Among 195 AIDS patients, 189 (96.9%) were farmers; 116 (59.5%) were men while 79 were women; 146 (74.9%) were between 20 - 50 years old; 180 (92.3%) were married. The major routes of HIV transmission were blood/plasma donation or transfusion (176 patients, 90.3%). CD(4)(+) T cell counts were between (1 - 1531) × 10(6) cells/L ((323.6 ± 14.8) × 10(6) cells/L), with 44 (26.5%) patients' CD(4)(+) T cell counts less than 200 × 10(6) cells/L. Of which, 154 patients (79.0%) had sexual partners. 86.8% (118 patients) consistently used condoms during the past 6 months, while for the last sexual act, 91.8% (123 patients) used condoms. For anti-HSV-1 status, there were about 164 patients (84.1%) were positive, and 26 (13.3%) were positive for anti-HSV-2. While, 14 (7.2%) were positive for both anti-HSV-1 and anti-HSV-2. Logistic regression analysis indicated that marital status were correlated with HSV-2 infection (OR = 7.41; 95%CI: 2.42 - 22.73; P < 0.01). No socio-demographic and sexual characteristics were identified to be correlated with HSV-1 infection.

CONCLUSIONA substantial proportion of AIDS patients in a rural county of Shanxi province of China were co-infected with HSV-1 and/or HSV-2. Marital status was correlated with HSV-2 infection.

Acquired Immunodeficiency Syndrome ; complications ; epidemiology ; virology ; Adolescent ; Adult ; Child ; China ; epidemiology ; Female ; Herpes Simplex ; complications ; epidemiology ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Logistic Models ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Rural Population ; Young Adult

This study was purposed to prepare and identify monoclonal antibodies (McAbs) against Homo sapiens hemoglobin alpha 2 (HBA2). Normal human fetal liver tissues were homogenized, and human liver nuclear proteins were isolated by centrifugation. The total human fetal liver nuclear proteins were used to immunize BALB/c mice for preparing McAbs by hybridoma technique. The McAbs specificity was identified by ELISA, Western blot, and immunohistochemistry. The antigen was identified by Uni-ZAP expression library screening. The results showed that one hybridoma cell line, AEE091, secreting specific McAb against HBA2 was established. The Ig subclass of this McAb was IgG1 (kappa). Data from immunohistochemistry assay showed that AEE091 could recognize human liver nuclear protein. Using AEE091 McAb, isolation of the protein antigen by IP revealed that AEE091 McAb could recognize 15 kD protein. Screening the Uni-ZAP XR pre-made liver cDNA library with AEE091 hybridoma cell supernatants demonstrated that AEE091 McAb specially reacted with HBA2. It is concluded that a hybridoma cell line stably secreting specific McAb against HBA2 is established. The specific McAb against HBA2 would be very useful for studying HBA2 function and screening thalassemia.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; Base Sequence ; Hemoglobin A2 ; immunology ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; alpha-Thalassemia ; immunology

OBJECTIVETo study the feasibility of vector-mediated RNA interference for HER-2-positive breast cancer therapy.

METHODSA plasmid vector capable of mediating HER-2 RNA interference was constructed, and HER-2-positive breast cancer cell line SKBR-3 was transfected with this constructed vector. The expression of HER-2 mRNA and protein was analyzed by RT-PCR and Western blotting, and the growth and apoptosis of SKBR-3 cells was analyzed after transfection.

RESULTSThe expressions of HER-2 mRNA and HER-2 protein was downregulated in response to vector-mediated HER-2 RNA interference, which also resulted in tumor cell growth inhibition and increased number apoptotic cells.

CONCLUSIONHER-2 is a good target for RNA interference and RNA interference targeting HER-2 can lead to HER-2 breast cancer cell apoptosis and growth inhibition.

Apoptosis ; Blotting, Western ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Genetic Vectors ; Humans ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Receptor, ErbB-2 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection