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.

Objective: To assess changes of liver function in HIV-positive children with/without HBV/HCV co-infection after 1 year of highly active antiretroviral therapy ( HARRT ) . Methods: Seventy-eight pediatric AIDS patients with HBV/HCV co-infection,19 pediatric AIDS patients with HBV co-infection and 44 pediatric AIDS patients without HBV/HCV co-infection who received HAART at least for 1 year were enrolled .HIV-1 viral load was quantitatively detected using a standardized reverse transcriptase-polymerase chain reaction assay , and blood cells were determined by three-color flow cytometry . Anti-HCV antibody and HBsAg was detected using an enzyme-linked immunosorbent technique , and ALT, AST and TBIL were detected by automatic biochemical analyzer .Results: After 1 year-HAART, the viral load was decreased to the lowest limit of detection in 90 .34% patients ( t=2 .61 , P<0 .01 ) , and CD4 +T cell counts were increased from 170 .187 ±132 .405/μl to 796 .014 ± 158 .491/μl ( t=3 .17 , P<0 .01 ) .The levels of ALT and AST were elevated ( t=2 .02 , P <0 .05 ) , while the ALT and AST levels in patients receiving nevirapine (NVP) based HAART increased from 18.28 ±13.74 U/L and 24.23 ±8.09 U/L to 55.35 ±22.40 U/L and 69.97 ±26.72 U/L, respectively(t=3.80,t=4.11;Ps<0 .01 ) .The increment of ALT and AST in NVP based HAART were significantly higher than that in the efavirenz based HAART (ALT:46.28 ±13.35 U/L vs 37.70 ±15.25 U/L and AST:19.53 ±7.23 U/L vs 1.25 ±0.21 U/L, respectively; t=4.53, t=5 .79;Ps<0 .01 ) , particularly in patients co-infected with HIV/HBV/HCV ( ALT:54 .32 ±22 .85 U/L vs 16 .89 ±14 .42 U/L and AST:41 .71 ±19 .26 U/L vs -3 .44 ±15.59 U/L, respectively; t=3.42, t=2.98, Ps<0.01).Conclusion: HARRT can repress HIV-1 replication effectively , but it also cause the damage of liver function , especially in patients with HBV and/or HCV co-infection.

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 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

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

Objective To study the genetic variations between measles vaccinc strain S191 and strains that circulated in Zhejiang province causing the epidemics during 1999 to 20 1 1.Methods Complete sequence of the nine Zhejiang measles strains were amplified by RT-PCR assay.Products were sequenced and the obtained sequences were aligned and analyzed with vaccine strains S191 and the major epidemic strains isolated in foreign countries.Results The homology of amino acid among the nine Zhcjiang strains were 98.77％-99.89％.The strains were not affected by positive selection and the variations on each gene were still in random drift.Compared to vaccine strain S191,there were 135 to 159 amino acid changes in Zhejiang measles virus,in which 113 points were common variable positions,resulting in mutations on five glycosylation sites.At the nucleotide level,the biggest differences between the Zhejiang strains and the vaccine strain S191 were found on N gcne,with the average divergent ratio as 5.5％,while the biggest one was P protein,in the amino acid level,with the average mutation rate as 7.7％.In addition,with the complete genome sequences,the genetic distance between Zhcjiang epidemic strains and vaccine strains was greater than the distances between epidenic strains of genotype D4,B3 and vaccine strains (t=-9.76,P＜0.05；t=-12.39,P＜0.05).Conclusion There were significant differcnccs found in the each of the genes between Zhejiang epidemic strains and the vaccine strain S191.The differences between the current vaccine strains and H genotypc epidemic strains were much larger than the differences between the vaccine and the forcign epidemic strains (genotype D4,B3).Therefore,wc should pay close attcntion to this trend,and to develop candidates for the dcvclopnent of vaccines,as early as possible.