To compare the hemodynamic effects of propofol, midazolam and etomidate during induction in sick children.Methods:Sixty sick children were randomly divided into three group s to receive intravenous propofol 2.5mg·kg-1(groupP),midazolam 0.2mg·kg-1(group M) or etomidate 0.3mg·kg-1(groupE), respectively. All hemodynamic data were recorded before administration, during intubation a nd 1,3,5,10 min after intubation. Results: The Bp and HR h ad no change in group Pand group Mduring whole procedure. In group E, the Bp had increased significantly at intubation to 1,3 min after intubation. Conclusion: propofol, midazolam and etomidate may be safely adminis trated for sick children during induction of anaesthesia.

Objective The construction of suicide plasmid vector could be used to make mutation of pgm gene which attenuates the virulent of Brucella melitensis strain 16, the research may lay a foundation for the development of novel live attenuated vaccines. Methods Sucrose sensitive gene as forward screening sign and fusion sequences of kanamycin resistance gene were constructed based on plasmid pucl9; pucS1.6K suicide plasmid vector was established by modifying pgm gene with fusion sequences of kanamycin resistance gene (insertion mutation); pgm gene mutation of Brucella melitensis strain 16 was obtained by electro transformation and mutation was confirmed by PCR amplification. Results The results showed that the identified Brucella melitensis strain 16 pgm gene was inactivated after insertion of kanamycin resistance gene, and the mutant pgm gene DNA fragment length was approximately 3525 bp, in line with expectations, Brucella pgm gene mutant melitensis strain 16 was successfully constructed. Conclusions The construction of suicide plasmid vector and precise mutation of Brucella melitensis strain 16 is successful, the study is not only provided an effective technology platform for constructing mutants of Brucella but also lays a foundation for the development of novel live attenuated vaccines.

Acute Disease ; Adolescent ; Coronary Angiography ; Coronary Sinus ; abnormalities ; diagnostic imaging ; Coronary Vessel Anomalies ; complications ; diagnostic imaging ; Electrocardiography ; Female ; Humans ; Myocardial Infarction ; diagnostic imaging ; etiology ; Tomography, X-Ray Computed

OBJECTIVEThe determination of antigenicity and immunogenicity of Leptospira interrogans genus-specific outer envelope proteins (OEPs) will offer evidence for developing universal leptospiral genetic engineering vaccine and detection kit.

METHODSIn this study, Ni-NTA affinity chromatography is used to purify the recombinant products rLipL21, rOmpL1/1, rOmpL1/2, rLipL32/1, rLipL32/2, rLipL41/1 and rLipL41/2 expressed by the major genotypes of four leptospiral OEPs of 15 serogroups. SDS-PAGE is applied to examine the expression and purity of the recombinant proteins. Rabbits are intracutaneously immunized with the recombinant proteins to obtain antisera. Microscope agglutination test (MAT) is used to measure the cross inmmunoagglutination titers of antisera. The OMPs of the reference standard strains belonging to 15 serogroups of L. interrogans in China and L. biflexa strain Patoc I are prepared using salt-denature method. By each of the antisera as the first antibody, Western blot assay is established to detect the natural expressions and immunoreactivity of the four OEPs.

RESULTSThe outputs of rLipL21, rLipL32/1, rLipL32/2, rLipL41/1l, rLipL41/2, rOmpL1/1 and rOmpL1/2 are 10%, 40%, 35%, 15%, 10%, 30% and 15%, respectively. Each the purified recombinant proteins shows a single fragment after SDS-PAGE. Each the rabbit antisera displays extensive cross immunoreactivity between the products expressed by different genotypes of the same gene and the MAT titers ranging from 1:2-1:128. All the four OEPs can be detectable in the OEPs preparations. However, LipL21 is found to exist only in L. interrrogans.

CONCLUSIONThe results of this study indicate that all the four OEPs are superficial genus-specific antigens of Leptospira which can be used as the candidate antigens of leptospiral universal vaccine and detection kit.

Animals ; Antibody Formation ; Antigens, Bacterial ; immunology ; Electrophoresis, Polyacrylamide Gel ; Genetic Engineering ; Immunization ; Leptospira interrogans ; classification ; immunology ; Membrane Proteins ; Rabbits ; Recombinant Proteins ; immunology ; Serotyping

OBJECTIVETo investigate the molecular mechanism of curcumin in reducing blood lipids by establishing gene expression system of human low density lipoprotein receptors (LDL-R) in Xenopus Laevis oocytes (XLO).

METHODSThe expression of LDL-R on cytomembrane was determined using immuno-fluorescent, ligand-fluorescent and immune colloidal gold techniques after human LDL-R containing p3.7 LDL plasmid was led into nucleus. And the expression of LDL-R gene in XLO was quantitatively determined by ELISA after being interfered with different concentrations of curcumin.

RESULTSThe human LDL-R gene could be expressed on XLO, which could be significantly enhanced by curcumin in a dose-dependent manner. Conclusion One of the paths of curcumin in reducing blood lipids and anti-atherosclerosis was improving LDL-R gene expression and increasing the LDL-cholesterol absorption of cells.

Animals ; Cells, Cultured ; Curcumin ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Gene Expression Regulation ; Humans ; Microinjections ; Oocytes ; cytology ; metabolism ; Receptors, LDL ; biosynthesis ; genetics ; Xenopus laevis

Abstract: Objective　To investigate the drug demand and related influencing factors of AIDS non-occupational post-exposure prophylaxis (nPEP) among men who have sex with men (MSM) in Wuhan, and to provide a scientific basis for the development of subsequent intervention policies for MSM. Methods　With the assistance of social organizations in Wuhan, MSM was recruited by the snowball method to carry out an online questionnaire survey to collect information on demographics, AIDS-related knowledge, high-risk behaviors, and the need for nPEP medication. The χ2 test and unconditional Logistic regression were used to analyze the related factors of the demand for nPEP medication. Results　A total of 308 valid subjects were included in this study, with predominantly 18-29 years old (78.57%, 242/308). The self-reported sexual orientation was mainly homosexuality (82.47%, 254/308), and the awareness rate of AIDS knowledge was high (89.29%, 275/308). Among the survey respondents, 35.06% (108/308) did not know the situation of HIV infection among MSM population in Wuhan; 55.19% (170/308) had two or more same-sex sexual partners in the last six months; 90.91% (280/308) had heard of nPEP before participating in this survey. After passing nPEP and informing the protective effect of nPEP, 59.42% (183/308) of them needed nPEP. After HIV exposure, 73.38% (226/308) were willing to spend money to buy nPEP drugs, and 88.64% (273/308) were willing to take nPEP drugs because of the known side effects. Logistic regression analysis showed that in the last six months, the needs for taking nPEP medication in those who had 2 or more same-sex sexual partners (OR=2.121, 95%CI: 1.329-3.386) and who had received peer education (OR=1.740, 95%CI: 1.088-2.781) were higher than those of those who had a same-sex sexual partner in the last six months and who had not received peer education. Conclusions　The MSM population in Wuhan has a great demand for nPEP drugs, and peer education is an important way to carry out nPEP publicity and promotion. At the same time, we should continue to strengthen warning publicity and education and behavioral intervention to reduce MSM risky sexual behaviors and reduce new HIV infections.

Eleven flavonol glycosides were isolated from the ethanol extract of Lysimachia clethroides by a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and reversed-phase HPLC. Their structures were identified as astragalin (1), isoquercitrin (2), isorhamnetin-3-O-β-D-glucopyranoside (3), quercetin-3-O-β-D-6"-acetylglucopyranoside (4), quercetin-7-O-β-D-glucopyranoside (5), prunin (6), 2-hydroxynaringin-5-O-β-D-glucopyranoside (7), kaempferol-3-O-rutinonoside (8), kaempferol-3-O-robinobioside (9), rutin (10) and kaempferol-3,7-di-O-β-D-glucopyranoside (11). Among them, compounds 4, 7 and 11 were obtained from the Lysimachia genus for the first time, while compounds 3, 5 and 9 were firstly reported from this plant. In the preliminary assays, compounds 2, 6 and 8 possessed significant inhibition against aldose reduc- tase, with IC50 values of 2.69, 1.00, 1.80 μmol · L(-1), respectively; none of compounds 1-11 exhibited obvious cytotoxic activity (IC50 > 10 μmol · L(-1)).
Drugs, Chinese Herbal ; chemistry ; Flavonols ; chemistry ; Glycosides ; chemistry ; Molecular Structure ; Primulaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo study regulation of Ca(2+)-activated K(+) channels (KCa) of mesenteric artery smooth muscle cell (SMC) from 21 old patients with essential hypertension (EH) by endothelin-1 (ET-1) and prostagl E(1) (PGE(1)).

METHODSMesenteric artery branch from EH was digested by enzyme. Patch clamp technique was used to pull cell-attached and inside-out patches on mesenteric artery SMC from EH and the normotensive patients respectively. The signal channel open probability (Po), open dwell-time (To) and close dwell-time (Tc), open channel number per patch were recorded. After adding Ca(2+) (10(-8) approximately 10(-6) mol/L), ET-1(2 approximately 8 x 10(9) mol/L) and PGE(1) (10, 20, 40, 100, 200, 400 nmol/L) to cytoplasm respectively. The parameters above were observed again.

RESULTSCompared to that of normotensive patients, the activities of KCa channels of patients with EH was higher. After adding Ca(2+) to cytoplasm,the Po of KCa channels in normotensive patients increased significantly. But it was few changes in EH group. KCa channels has dual reaction to ET-1 in normotensive patients. We have found no statistics difference when ET-1 present on KCa channels of EH cases. Whereas PGE(1) can affect KCa channels current and channels kinetic significantly in side-out patches. The Po of KCa channels increased. The To protracted and the Tc curtailed in EH.

CONCLUSIONSThe activities of KCa channels of patients with EH increased significantly. but the sensitive to Ca(2+) decreased. ET-1 were few effect to KCa channels. The PGE(1) can activated KCa channels of patients with EH.

Aged ; Alprostadil ; pharmacology ; Cells, Cultured ; Endothelin-1 ; pharmacology ; Female ; Humans ; Hypertension ; metabolism ; physiopathology ; In Vitro Techniques ; Male ; Mesenteric Arteries ; cytology ; Middle Aged ; Muscle, Smooth ; metabolism ; physiopathology ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; drug effects ; metabolism

Animals ; Antibodies, Bacterial ; blood ; Bacterial Proteins ; genetics ; immunology ; Brucella Vaccine ; immunology ; Brucella abortus ; immunology ; Female ; Immunization ; Interferon-gamma ; biosynthesis ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Periplasmic Binding Proteins ; genetics ; immunology ; Ribosomal Proteins ; genetics ; immunology ; Vaccines, DNA ; immunology

OBJECTIVETo study the specific cellular immune response induced by a recombinant HBV DNA vaccine and evaluate the potential therapeutic effect of this vaccine against HBV.

METHODSA series of HBV epitopes were screened and the optimal combinations of these gene fragments identified to construct the target gene HS which incorporated appropriate flanking sequences. HS was inserted in pVAX1 to construct the recombinant plasmid PVAX-HS. Generation of specific cytotoxic T lymphocytes (CTLs) induced by PVAX-HS in HLA-A2 mice was observed by 51Cr assay and the activity of the CTLs evaluated using ELISPOT assay.

RESULTSPVAX-HS was constructed successfully, which induced specific CTLs with strong activities after immunization in mice as compared with the control group that yielded negative results.

CONCLUSIONPVAX-HS DNA immunization can induced specific cellular immune response in mice, suggesting the potential therapeutic effect of this HBV vaccine.

Animals ; Hepatitis B Vaccines ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Humans ; Immunization ; methods ; Interferon-gamma ; biosynthesis ; Mice ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; Vaccines, DNA ; genetics ; immunology