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.

OBJECTIVETo understand the pollution rates of vibrio cholera (V. cholera) in different seafood, aquatic products and their circulatory processes, so as to help making measures for cholera control and prevention.

METHODSDifferent seafood, aquatic products and breed water specimen collected from 12 provinces of China were tested from July to September in 2005.

RESULTA total of 12 104 samples of seafood and aquatic products were tested and the average pollution rate of vibrio cholera was 0.52%. The positive isolate rate of turtle sample (1.72%) was the highest among all samples. The second higher isolated rate was 1.14% in water specimen of turtle breed pool. The positive rate of bullfrog was 0.50%. The percentage of toxin strains was 47.54% and 79.31% of them were isolated from turtle and water samples of turtle breed pool. The important sector of the pollution of vibrio cholera was in turtle breed pool (2.38%).

CONCLUSIONThe average pollution rate of vibrio cholera in seafood and aquatic products in 12 provinces of China was low. It should be very necessary to supervise the sanitation in turtle breed for controlling and preventing the vibrio cholera.

Animals ; China ; Female ; Fishes ; microbiology ; Food Contamination ; analysis ; prevention & control ; statistics & numerical data ; Male ; Seafood ; microbiology ; Seawater ; analysis ; Turtles ; microbiology ; Vibrio cholerae ; isolation & purification

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

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

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