Acquired Immunodeficiency Syndrome ; immunology ; Adolescent ; Chemokines ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Male

Foreign Bodies ; Humans ; Male ; Middle Aged ; Skull Base

Objective To construct the bait expression plasmid pGBKT7-GR of glucocorticoid receptor(GR)binding domain.Methods The fragments of GR binding domain was amplified by RT-PCR,and then was cloned into pMD18-T.After being verified by sequencing,it was subcloned into the bait expression vector pGBKT7.Then the bait vector pGBKT7-GR was transformed into AH109 yeast cells and the expression of the bait protein was analyzed by Western blot.Toxicity and self-activation of the bait protein were detected.Results GR binding domain was amplified and cloned into pMD18-T and pGBKT7 successfully.The bait vector was transformed into AH109 yeast cells successfully,without toxicity or self-activation.The expression of the bait protein was confirmed by Western blot.Conclusion The successful construction of bait expression vector of glucocorticoid receptor binding domain lays the foundation for constructing small molecule ligand yeast three-hybrid system.

Acute promyelocytic leukemia (APL) is characterized by the generation of the prototypic promyelocytic leukemia-retinoicacid receptor alpha (PML-RARα), an oncogenic fusion protein due to chromosomal translocation. In a human myeloid cell line,PML-RARα is cleaved by neutrophil elastase (NE) to produce the mutational PML [nuclear localization signal (NLS) deletion andRARα (NLS-RARα, containing NLS of PML), both of which may play an important role in APL pathogenesis. The yeast two-hybridtechnique was used to screen the intracellular proteins interacting with NLS-RARα, which may be involved in NLS-RARα signaling. The NLS-RARα coding sequence was amplified by polymerase chain reaction method and was cloned into the bait plasmid pGBKT7vector, which, after the confirmation by sequencing, was transformed into yeast AH109 and the subsequent expression of bait plasmidwas proved by Western-blot. The transformed yeast AH109 was mated with yeast Y187 (containing leukemia cDNA library plasmidspACT2) in medium. Diploid yeast was plated on synthetic dropout nutrient medium containing X-α-gal for screening. After beingreintroduced into yeast AH109 and sequenced to verify the expression of ORF, eight positive colonies were obtained, among whichonecontaining JTV-1 was cloned. The interaction between NLS-RARα and JTV-1 was further supported by indirect immunofluorescence,GST pull-down and co-immunoprecipitation, respectively. These findings brought some new clues for the further exploration ofNLS-RARα signaling to APL.

After estrogen receptor?(ER?)in MC,63 cell was knocked down by RNA interference, expression of osteoprotegerin(OPG)induced by 17?-estradiol was assayed by RT-PCR.17?-estradiol with various concentration obviously upregulated the expression of OPG mRNA of MG63 cell with the maxima]effect at the concentration 10~(-7)mol/L,which was not inhibited by suramin(G protein inhibitor).The present result suggested that ER?was not involved in regulation of OPG mRNA expression in MC,63 cell by 17?-estradiol.

OBJECTIVETo investigate the modulatory effects of morphine on the susceptibility to pentylenetetrazole-induced seizures, and the involvement of endogenous histamine in this process.

METHODSBoth the wild-type (WT) mice and histidine decarboxylase (a key enzyme for histamine biosynthesis) deficient (HDC-KO) mice were subcutaneously injected with different doses of morphine, and 1 hour later the pentylenetetrazole solution (1.5 %) was infused into the tail vein at a constant rate of 0.3 ml/min. The minimal dose of pentylenetetrazole (mg/kg) needed to induce myoclonic jerks and clonus convulsion was recorded as the thresholds of seizures.

RESULTIn WT mice, morphine dose-dependently decreased the thresholds of both myoclonic jerks and clonus convulsion. In HDC-KO mice, morphine at 10 mg/kg only significantly decreased the threshold of myoclonic jerks from (38.6 +/-2.9)mg/kg to (32.5 +/-0.7)mg/kg, but had no significant effect on the threshold of clonus convulsion [from (51.8 +/-2.1)mg/kg to (47.6 +/-1.2)mg/kg]. In addition, the value of decreased myoclonic jerks (15.8 +/-1.4)% and clonus convulsion (8.3 +/-0.9)% thresholds were much lower in HDC-KO mice than in WT mice [(26.1 +/-2.5)% and (20.8 +/-2.4)%, respectively].

CONCLUSIONMorphine can decrease the thresholds of pentylenetetrazole in induction of seizure, and the endogenous histamine may be involved in this process.

Animals ; Disease Susceptibility ; chemically induced ; metabolism ; physiopathology ; Dose-Response Relationship, Drug ; Histamine ; metabolism ; physiology ; Histidine Decarboxylase ; genetics ; metabolism ; Male ; Mice ; Mice, Knockout ; Morphine ; pharmacology ; Myoclonus ; chemically induced ; metabolism ; physiopathology ; Narcotics ; pharmacology ; Pentylenetetrazole ; Random Allocation ; Seizures ; chemically induced ; genetics ; physiopathology ; Sensory Thresholds ; drug effects

OBJECTIVETo compare the long-term efficacy of procedure for prolapse and hemorrhoids (PPH) and Milligan-Morgan hemorrhoidectomy (MMH) in the treatment of III and IV degree internal hemorrhoids.

METHODSOne hundred patients were randomly divided into two groups and received PPH (n=42) and MMH (n=58) respectively. After two years, the efficacy, complications and function of defecation were compared.

RESULTSTwo years after operation, the morbidities of hydrorrhea (2.38% vs 20.69%, P=0.007), dermal neoplasm formation (9.52% vs 25.86%, P=0.040) and narrowing in the caliber of the stools (2.38% vs 18.97%, P=0.027) were significantly lower in PPH group than those in MMH group (P<0.05). The morbidities of overall complication (9.52% vs 25.86%, P=0.040) and overall abnormal function of defecation (9.52% vs 29.31%, P=0.017) were lower in PPH group than those in MMH group (P<0.05). However, there were no significant differences of the morbidity of relapse (14.29% vs 10.34%, P=0.549), patient satisfactory degree (92.86% vs 87.93%, P=0.636) and overall symptom recurrence rate (19.05% vs 25.86%, P=0.424) between the two groups.

CONCLUSIONSLong-term efficacies of procedure for prolapse and hemorrhoids and Milligan-Morgan haemorrhoidectomy in the treatment of III and IV degree internal hemorrhoids are similar. PPH has better safety, less complications and less effect on abnormal function of defecation compared with MMH.

Adolescent ; Adult ; Aged ; Female ; Hemorrhoids ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Mucous Membrane ; surgery ; Prolapse ; Sutures ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effect of plant growth substances on induction and culture of callus from Rhodiola quadrifida and also to analyze salidroside contents in the callus.

METHODThe optimum combination of plant growth substances in MS solid medium for induction and culture of callus was established using orthogonal design. The contents of salidroside was analyzed by HPLC.

RESULTMS medium containing 2,4-D 1 mg x L(-1), NAA 2 mg x L(-1), 6-BA 0.5 mg x L(-1) and KT 0.1 mg x L(-1) could induce the callus from R. quadrifida most effectively;the induction rate was 83.3%. The optimized combination of plant growth substances for callus subculture was 2,4-D 1 mg x L(-1), 6-BA 0.1 mg x L(-1) and KT 0.5 mg x L(-1). The dry weight could reach 11.77 g x L(-1) when the callus was cultured in the optimum medium for 30 d and salidroside content was 0.28%.

CONCLUSIONThe quantities of plant growth substances required for induction and culture of callus are different in R. quadrifida. The callus could produce salidroside.

Culture Media ; Glucosides ; metabolism ; Phenols ; metabolism ; Plant Growth Regulators ; pharmacology ; Plant Stems ; growth & development ; metabolism ; Plants, Medicinal ; growth & development ; metabolism ; Rhodiola ; growth & development ; metabolism ; Tissue Culture Techniques ; methods

AIMTo study the dissociation pathways of aminoglycoside antibiotics.

METHODSIn positive mode, eight aminoglycoside antibiotics were elucidated by use of electrospray ion trap mass spectrometry in the multi-stage MS full scan mode.

RESULTSIt was demonstrated that the eight aminoglycoside antibiotics gave abundant product ions at m/z 322 (gentamicin, micronomicin and sisomicin), m/z 350 (etimicin, netilmicin and vetilmicin) and m/z 324 (kanamycin and tobramycin) by loss of the C-ring (amino-alpha-D-glucopyranose) in MS2 full scan mode. In MS3 full scan mode, the prominent fragmentation ions at m/z 163 as well as m/z 191 were formed from the fragmentation ions at m/z 322, m/z 350 and m/z 324 by loss of the A-ring (amino-alpha-D-glucopyranose), separately, while the characteristic fragmentation ions at m/z 160 as well as m/z 162 were formed from m/z 322, m/z 350 and m/z 324 by loss of the B-ring (2-deoxy-D-streptamine), separately.

CONCLUSIONThe structural information was obtained via collision-activated dissociation and these characteristics are applicable to the structural elucidation and quantitative analysis of aminoglycoside compounds.

Aminoglycosides ; analysis ; chemistry ; Gentamicins ; analysis ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tobramycin ; analysis ; chemistry

The aim of this paper is to report the synthesis of the mPEG-PCL-g-PEI copolymers as small interfering RNA (siRNA) delivery vector, and exploration of the siRNA delivery potential of mPEG-PCL-g-PEI in vitro. The diblock copolymers mPEG-PCL-OH was prepared through the ring-opening polymerization. Then, the hydroxyl terminal (-OH) of mPEG-PCL-OH was chemically converted into the carboxy (-COOH) and N-hydroxysuccinimide (NHS) in turn to prepare mPEG-PCL-NHS. The branched PEI was reacted with mPEG-PCL-NHS to synthesize the ternary copolymers mPEG-PCL-g-PEI. The structure of mPEG-PCL-g-PEI copolymers was characterized with Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR) and gel permeation chromatography (GPC). The mPEG-PCL-g-PEI/siRNA nanoparticles were prepared by complex coacervation, and the nanoparticles size and zeta potential were determined, separately. The cytotoxicities of mPEG-PCL-g-PEI/siRNA nanoparticles and PEI/siRNA nanoparticles were compared through cells MTT assays in vitro. The inhibition efficiencies of firefly luciferase gene expression by mPEG-PCL-g-PEI/ siRNA nanoparticle at various N/P ratios were investigated through cell transfection in vitro. The experimental results suggested that the ternary (mPEG5k-PCL(1.2k))1.4-g-PEI(10k) copolymers were successfully synthesized. (mPEG(5k)-PCL(1.2k))1.4-g-PEI(10k) could condense siRNA into nanoparticles (50-200 nm) with positive zeta potential. MTT assay results showed that the cytotoxicity of (mPEG(5k)-PCL(1.2k))1.4-g-PEI(10k)/siRNA nanoparticles was significantly lower than that of PEI(10k)/siRNA nanoparticles (P < 0.05). The expression of firefly luciferase gene could be significantly down-regulated at a range of N/P ratio from 50 to 150 (P < 0.01), and maximally inhibited at the N/P ratio of 125. The mPEG-PCL-g-PEI polymers could delivery siRNA into cells to inhibit the expression of target gene with very low cytotoxicity, which suggested that mPEG-PCL-g-PEI could serve as a new type of siRNA delivery vector.
Cell Line, Tumor ; Cell Survival ; Drug Carriers ; Genes, Reporter ; Genetic Vectors ; Humans ; Luciferases ; metabolism ; Molecular Weight ; Nanoparticles ; Particle Size ; Polyesters ; chemistry ; Polyethylene Glycols ; chemistry ; Polyethyleneimine ; chemistry ; Polymers ; chemistry ; RNA, Small Interfering ; administration & dosage ; genetics ; metabolism ; Transfection