Adolescent ; Adult ; Female ; Health Care Surveys ; Health Status ; Humans ; Male ; Medical Waste Disposal ; Middle Aged ; Occupational Health ; Personnel, Hospital ; Young Adult

OBJECTIVETo economically obtain the Abeta peptide for Alzheimer's disease (AD) research by expressing the Abeta peptide fused with the maltose binding protein (MBP) possessing high solubility in E.coli.

METHODSThe cDNA-coding sequence of Abeta peptide was modified by the addition of a BamH I site at the 5' end and a Hind III site at the 3' end using PCR. The modified sequence was ligated into the maltose-binding protein (MBP) fusion expression vector pMAL-c2 containing an thrombin cleavage site, which was transformed into competent E.coli DH5alpha cells. After identification of the single clones by PCR and DNA sequencing, the recombinant plasmid was transformed into E.coli TB1 and induced to express MBP-Abeta fusion protein. The expressed fusion protein was purified using amylose resin column and identified by SDS-PAGE and Western blotting.

RESULTSThe result of DNA sequencing verified the consistency between the inserted sequence and Abeta (1-42) sequence. SDS-PAGE electrophoresis showed that MBP-Abeta fusion protein was highly expressed in E.coli TB1, and Western blotting demonstrated that the purified fusion protein and the separated Abeta peptide could be recognized by specific anti-Abeta (22-35) antibody.

CONCLUSIONMBP-Abeta fusion protein highly expressed in E. coli TB1 cells with enhanced solubility and the separated Abeta peptide with good immunogenicity obtained may lend support to AD research.

Alzheimer Disease ; genetics ; Amyloid beta-Peptides ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Maltose-Binding Proteins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

Chemical constituents of ethyl acetate extract of Illicium burmanicum were isolated and purified by various chromatographic methods,including Silica gel, Sephadex LH-20, C18 reverse-phased silica gel, Preparative TLC and Preparative HPLC. Their structures were identified by spectral analysis including NMR and MS data. Fourteen compounds were separated from I. burmanicum and their structures were identified as 7S,8R-erythro-4,7,9,9'-tetrahydroxy-3,3'-dimethoxy-8-O-4'-neolignan (1), 7R,8R-threo-4,7, 9,9'-tetrahydroxy-3,3 '-dimethoxy-8-O-4'-neolignan(2) ,polystachyol(3), (-) -massoniresinol(4), angustanoic acid F (5), trans-sobrerol(6), (3S,6R) -6,7-dihydroxy-6,7-dihydrolinalool (7), (3S, 6S) -6,7-dihydroxy-6,7-dihydrolinalool (8), 2,6-dimethoxy-4-allyl-phenol (9), 3,5-dihydroxy4-hydroxy benzaldehyde (10), 3-hydroxy4-methoxybenzaldehyde (11), methyl vanillate (12), shikimic acid ethylester (13) and beta-sitosrerol (14). Except compound 14, the rest thirteen compounds were separated from this plant for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Illicium ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Through study on the correlation between Atractylodis macrocephala lactones ingredient content and topographic factors, we researched regionalization from topography of five main producing provinces of the country, in order to provide a scientific basis for A. macrocephala reasonable cultivation. By sampling from 40 origins of five main producing provinces of the country, the variation of A. macrocephala lactones ingredient content in different conditions of topographic factors and the effect of altitude, slope and aspect was analyzed by SPSS. Then according to the relationship between A. macrocephala lactones ingredient content and topographic factors, the ecological suitability regionalization was conducted by using ArcGIS based on topographic factors. It is suitable for growth of A. macrocephala in the hilly and mountainous areas of southern whose A. macrocephala lactones ingredient content is in high levels. It is unsuitable for growth of A. macrocephala in Northern plain areas, but we can cultivate A. macrocephala in the hilly and mountainous areas of Northern. The most suitable topographic condition for cultivation of A. macrocephala : altitude 200 meters above, slope 3.00-4.99 degrees.
Altitude ; Atractylodes ; chemistry ; growth & development ; China ; Conservation of Natural Resources ; Drugs, Chinese Herbal ; analysis ; Ecosystem

OBJECTIVETo investigate the effect and the mechanism of Shenfu injection (SFI) on the resuscitation from general anesthesia.

METHODSForty patients who received selective abdominal surgery with general anesthesia for 3-4 hrs and ASA grade I-II were divided into two groups, the trial group and the control group, 20 patients in each group. After being sent into the postanesthesia care unit (PACU), the trial group was treated with intravenous dripping of SFI 1.0 ml/kg and the control group was treated with intravenous dripping of equal volume of normal saline. All patients were observed in double blindly manner, the self ventilation recovery time, extubation time, the time of leaving PACU and their Glasgow coma scale (GCS) were recorded and compared. 2 ml of peripheral venous blood were taken to determine the plasma beta-endorphin (beta-EP) content at the time points of before (T1), 5 min (T2), 15 min (T3) and 30 min (T4) after dripping.

RESULTSThe self ventilation recovery time, extubation time and time of leaving PACU in the trial group were all shorter than those in the control group (P < 0.01), the GCS in the trial group was better than that in the control group (P < 0.01). The plasma content of beta-EP raised gradually along the recovering of patients consciousness, as compared with the content before dripping (T1), it showed insignificant difference at time point T2 but significant difference at T3 and T4 comparison at the corresponding time point showed that the content at T1 and T2 were similar in the two groups (P > 0.05), but at T3 and T4, the content was higher in the trial group than that in the control group respectively (P < 0.01).

CONCLUSIONSFI could accelerate the resuscitation after general anesthesia, the mechanism may be related with its action in raising plasma beta-EP level.

Adolescent ; Adult ; Aged ; Anesthesia Recovery Period ; Anesthesia, General ; Colonic Neoplasms ; surgery ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gastrectomy ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; beta-Endorphin ; blood

OBJECTIVETo identify the IgE-binding epitopes in the allergen Der p 1 of main house dust mites, which can be recognized by the specific IgE in the sera from allergic individuals, and obtain a hypoallergen derived from the T-B epitope fused peptide for potential use in specific immunotherapy (SIT).

METHODSThirty-one peptides containing 15 amino acids each, which covered the full 222 amino acids of Der p 1 protein sequence, were synthesized on the cellulous membrane by solid-phase peptide (SPOTs) synthesis, with 8 overlapping amino acids between every two neighboring peptides. The membrane bearing the spots of the synthesized peptides were incubated with the allergic serum pools consisting of the sera from 5 allergic individuals. The membrane was then probed with HRP-conjugated anti-human IgE, followed by enhanced chemiluminescence (ECL) for visualization and gray scale analysis of the positive peptide spots.

RESULTSThree strong IgE-binding epitopes were identified in the amino acid sequence of Der p 1 molecule, namely Ep1 (amino acids 85-99), Ep2 (amino acids 106-120) and Ep3 (amino acids 190-204).

CONCLUSIONThe 3 IgE-binding epitopes (B cell epitopes) identified in Der p 1 confirm the presence of linear epitopes in Der p 1, suggesting the possibility of constructing T/B epitope-fused hypoallergens.

Amino Acid Sequence ; Animals ; Antigens, Dermatophagoides ; immunology ; Arthropod Proteins ; immunology ; Binding Sites, Antibody ; Cysteine Endopeptidases ; immunology ; Epitopes ; immunology ; Immunoglobulin E ; immunology ; Lymphokines ; immunology ; Mites ; immunology ; Molecular Sequence Data

OBJECTIVETo investigate the displacement, the position of rotation center and the stress distribution of PDL under different loading force system (Moment/Force, M/F) by simulating clinical loading force system.

METHODSA three-dimensional finite element model of upper central incisor, which consists of 945 isoparametric elements and 1,245 nodes was developed. The displacement, the position of the rotation center and the stress distribution of PDL were analyzed under 13 types of loading force system.

RESULTS1. Different force system led to different types of tooth movement. When M/F= -9.15:1, -10.30 - -10.50:1 and -10.90:1, it brought the result of controlled tipping movement, the bodily movement and the root movement; 2. The graph of the center rotation was a hyperbolic asymptotic line: Mx/Fy = -10.50 (horizontal axis) and L = 6.75 (vertical axis). Moreover, a little change of M/F between -9.15 and -10.90 led to apparent change of the position of rotation center. 3. The maximum strain and stress during the tipping movement were 1.47 x 10(-2) MPa and -2.81 x 10(-2) MPa, and during the bodily movement the results were 1.10 x 10(-2) MPa and -1.86 x 10(-2) MPa, while during the root movement were 0.96 x 10(-2) MPa and -2.58 x 10(-2) MPa.

CONCLUSION1 . Different force system (M/F) leads to different types of tooth movement. 2. It is necessary to adjust the force system accurately to obtain prescient tooth movement, especially when M/F changes between -9.15:1 and -10.90:1. 3. This study suggested that the tooth movement style and the force system (M/F) should be controlled to protect the periodontal tissue.

Dental Stress Analysis ; Finite Element Analysis ; Humans ; Incisor ; physiology ; Models, Biological ; Periodontal Ligament ; physiology ; Tooth Movement Techniques

We have isolated a strain C611 that used methane as the sole carbon sources for growth from paddy soil in Taiyuan of Shanxi province. Based on the physiological characteristics and 16S rDNA sequence analysis, we identified the strain as Klebsiella sp.. We used statistic-based experimental design (RSM) to optimize the culture conditions for C611 strain. The optimum conditions were as follows: temperature of 24.4 degrees C, inoculum volume of 6.7% and methane content of 25%. We studied the response time and the relationship between consumption of dissolved oxygen and methane gas contents with PVA-H3BO3 immobilized cell of C611 using electrochemical method. The response time was no more than 100 s of this reaction system, and the linear range of detection of methane content was from 0 to 10%. The standard gas sample 3% methane was measured by this method with the mean content value of 3.09%, RSD of 3.48%, and the relative error of 3%. Hence, it has the potential in developing biosensor for methane.
Environmental Monitoring ; methods ; Klebsiella ; isolation & purification ; metabolism ; Methane ; analysis ; metabolism ; Soil Microbiology

Alkaloids ; pharmacology ; Animals ; Calcium ; metabolism ; Creatine Kinase ; blood ; Creatine Kinase, MB Form ; Drugs, Chinese Herbal ; pharmacology ; Female ; Ginsenosides ; pharmacology ; Injections ; Isoenzymes ; blood ; Male ; Malondialdehyde ; analysis ; Mitochondria, Heart ; drug effects ; pathology ; Myocardial Reperfusion Injury ; drug therapy ; pathology ; Myocardium ; ultrastructure ; Protective Agents ; pharmacology ; Rabbits ; Tetrahydroisoquinolines ; pharmacology

In order to facilitate the detection of newly emerging or rare viral infectious diseases, a negative-strand RNA virus-severe fever with thrombocytopenia syndrome bunyavirus, and a positive-strand RNA virus-dengue virus, were used to investigate RNA viral genome unspecific amplification by multiple displacement amplification technique from clinical samples. Series of 10-fold diluted purified viral RNA were utilized as analog samples with different pathogen loads, after a series of reactions were sequentially processed, single-strand cDNA, double-strand cDNA, double-strand cDNA treated with ligation without or with supplemental RNA were generated, then a Phi29 DNA polymerase depended isothermal amplification was employed, and finally the target gene copies were detected by real time PCR assays to evaluate the amplification efficiencies of various methods. The results showed that multiple displacement amplification effects of single-strand or double-strand cDNA templates were limited, while the fold increases of double-strand cDNA templates treated with ligation could be up to 6 X 10(3), even 2 X 10(5) when supplemental RNA existed, and better results were obtained when viral RNA loads were lower. A RNA viral genome amplification system using multiple displacement amplification technique was established in this study and effective amplification of RNA viral genome with low load was achieved, which could provide a tool to synthesize adequate viral genome for multiplex pathogens detection.
Bunyaviridae Infections ; diagnosis ; virology ; Cell Line ; DNA Ligases ; metabolism ; DNA, Complementary ; analysis ; genetics ; DNA-Directed DNA Polymerase ; metabolism ; Dengue ; diagnosis ; virology ; Dengue Virus ; genetics ; isolation & purification ; Genome, Viral ; genetics ; Humans ; Phlebovirus ; genetics ; isolation & purification ; RNA, Viral ; analysis ; genetics ; Reference Standards ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Load