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

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.

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

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 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

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 analyze the impact of diabetes mellitus on the clinicopathological factors and prognosis of patients with colorectal cancer.

METHODSA total of 599 patients with colorectal cancer treated between January 2000 and June 2007 were collected retrospectively. The patients were divided into diabetes mellitus (DM) group and non-diabetes mellitus (NDM) group. The pathologic factors data was compared between the two groups, and the Logistic multivariable analysis was performed. The Cox regression model analysis of prognosis data was applied in 402 patients who underwent radical surgery without preoperative neoadjuvant therapy.

RESULTSA total of 58 cases (9.7%) developed diabetes mellitus. Significant differences was found in the body-weight, age, hypertension between the two groups (P < 0.05), while no significant differences in the pathologic factors, such as tumor differentiation, invasion depth, lymph node involvement, TNM stage and lymphovascular invasion was found between the two groups (P > 0.05). There was no significant correlation between diabetes mellitus and the pathologic factors on the Logistic analysis (P > 0.05). Among the patients underwent radical surgery directly, neither disease progression curve (P = 0.521) nor overall survival curve (P = 0.909) presented significant differences between the two groups. It's not shown that diabetes mellitus was significantly associated with the prognosis of patients with colorectal cancer by using Cox regression analysis (P = 0.991).

CONCLUSIONSDiabetes mellitus does not significantly influence the clinicopathological factors and the prognosis of colorectal cancer in patients receiving radical surgery, and it requires more investigation.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; complications ; pathology ; Diabetes Mellitus ; Female ; Follow-Up Studies ; Humans ; Logistic Models ; Male ; Middle Aged ; Prognosis ; Retrospective Studies

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