Bel1, a transactivator of prototype foamy virus (PFV), plays pivotal roles in the replication of PFV. Previous studies have shown that Bel1 bears a nuclear localization signal (NLS), but its amino acid sequence remains unclear and the corresponding importins have not been identified. In this report, we inserted various fragments of Bel1 into an EGFP-GST fusion protein and investigated their subcellular localization by fluorescence microscopy. We found that the 215PRQKRPR221 fragment could direct nuclear localization, which accords with the consensus sequence K(K/R)X(K/R) of monopartite NLS. Point mutation experiments revealed that K218, R219, and R221 are essential for the nuclear localization of Bel1. The results of the GST-pulldown showed that the Bel1 fragment with residues 215-223, which bears the NLS, interacts with KPNA1, KPNA6, and KPNA7. This result suggests that KPNA1, KPNA6, and KPNA7 maybe involved in Bel1 nuclear translocation.
Cell Line ; Cell Nucleus ; genetics ; metabolism ; virology ; Humans ; Nuclear Localization Signals ; genetics ; metabolism ; Protein Binding ; Protein Transport ; Retroviridae Infections ; genetics ; metabolism ; virology ; Retroviridae Proteins ; chemistry ; genetics ; metabolism ; Spumavirus ; chemistry ; genetics ; physiology ; Trans-Activators ; chemistry ; genetics ; metabolism ; alpha Karyopherins ; genetics ; metabolism

Objective To evaluate the value of contrast-enhanced uhrasound(CEUS)for liver fibrosis.Methods Ninty-two chronic hepatitis B patients with histology diagnosis and fifteen health control were detected by CEUS,dynamic images and the time-intensity curve were analyed.Results Hepatic veinartery transit(HV-ATT)became shorter in the early hepatic cirrhosis patients [(7.5±2.1)s]compared to the control[(11.8±2.7)s].However,there was no difference between the fibrosis patients [(13.0±2.2)s]and the control.Furthermore,HV-ATT of the fibrosis patients with S1 to S3 stage were(11.7±2.7)s(S1),(11.8±3.0)S(S2),(11.8±2.3)s(S3)without significant difference.Conclusions CEUS is valuable in diagnosing earlier hepatic cirrhosis,but not fibrosis.

OBJECTIVEIn contrast to CD(4)(+) helper T-lymphocytes (T(H)), little is known about the transcriptional regulation of CD(8)(+) cytotoxic T-lymphocytes (Tc) and its role in the pathogenesis of asthma is unclear. This study was conducted to investigate the effect of T-bet and GATA-3 mRNA expression on profiles of type 1 and type 2 cytotoxic T lymphocytes in asthmatic children.

METHODTotally 38 asthmatic children, including acute attack group composed of 20 cases (age 3 - 13 years, mean 6.2 +/- 2.9), remission group with 18 cases (age 3 - 12 years, mean 6.1 +/- 2.5) and 20 healthy control children (age 3 - 12, 6.9 +/- 2.7) were recruited in this study from Sep. 2005 to Mar. 2006. The mRNA expression of T-bet and GATA-3 in the peripheral blood mononuclear cells were detected by using semi-quantitative PCR and Tc1, Tc2 cell numbers by flow cytometry analysis system.

RESULTT-bet mRNA in asthmatic children was lower than that in control group and lower in attack stage than in remission stage (0.14 +/- 0.04, 0.21 +/- 0.03, 0.28 +/- 0.03, P < 0.05). In contrast, GATA-3 mRNA was higher in asthmatic children than in control group and higher in attack stage than in remission stage (0.49 +/- 0.09, 0.44 +/- 0.08, 0.37 +/- 0.04, P < 0.05). It was shown that Tc1 percentage was lower in asthmatic children than those of control group and lower in attack stage than those of remission stage (6.6 +/- 2.4, 14.2 +/- 4.3, 31.2 +/- 3.8, P < 0.05). Tc2 percentage in asthmatic children was higher than that of control group and higher in attack stage than that of remission stage (10.0 +/- 4.2, 5.4 +/- 2.2, 3.5 +/- 1.1, P < 0.05). Spearman correlation analysis revealed that T-bet mRNA was positively correlated with Tc1 percentage (r = 0.704) and negatively correlated with Tc2 percentage (r = -0.629). GATA3 mRNA was negatively correlated with Tc1 percentage (r = -0.612) and positively correlated with Tc2 percentage (r = 0.673). The T-bet/GATA-3 mRNA ratio was positively correlated with Tc1 percentage (r = 0.731) and Tc1/Tc2 (r = 0.773), while negatively correlated with Tc2 percentage (r = -0.642).

CONCLUSIONThe imbalance of T-bet/GATA-3 mRNA expression is closely correlated with skewed Tc2 dominance in asthmatic children.

Adolescent ; Asthma ; genetics ; immunology ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Female ; GATA3 Transcription Factor ; genetics ; metabolism ; Humans ; Male ; RNA, Messenger ; genetics ; metabolism ; T-Box Domain Proteins ; genetics ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo study the survival and growth of Vibrio cholerae inside the Acanthamoeba polyphage.

METHODSSurvival and growth of Vibro cholerae O139, co-cultured with Acanthamoeba polyphaga, was observed inside the trophozoites and cysts, using Gram stain and electron microscope.

RESULTSViable O139 was observed inside the amoebal vacuoles in 24 hours. Vacuoles were filled with more bacteria along with the longer period of co-culture. The process of O139 infection with Amoebae would include uptake, formation of O139 vacuole, multiplication, trophozoites lysed and expel under electron microscopy. Some infected trophozoites could subsequently encyst and the surviving O139 could locate in the vesicles inside the cysts.

CONCLUSIONO139 might survive and multiply in the trophozoites and reside inside the cysts of Amoebae, suggesting that Acanthamoebae might serve as one of the environmental hosts of Vibro cholerae.

Acanthamoeba ; growth & development ; microbiology ; ultrastructure ; Animals ; Coculture Techniques ; Colony Count, Microbial ; Culture Media ; Vibrio cholerae O139 ; growth & development ; ultrastructure ; Water ; parasitology

OBJECTIVETo ascertain the genetic characterization and genotype of measles viruses isolated in Shanghai region, in 2005.

METHODSMeasles virus was isolated from throat swab specimens collected from suspected measles cases and 450 bp fragment of C terminus of nucleprotein (N) gene was amplified by RT-PCR. Sequence analysis was conducted to ascertain the genotype and to compare the difference of nucleotide with other measles virus strain published in GenBank.

RESULTS4 measles viruses were isolated from 10 throat swab specimens, and the sequence analysis indicated that they belonged to H1 genotype. The homogeneity of 450 nucleotides in the C terminal of the N gene was at 98%-98.2% as compared to H1 genotype (China93-7). They differed from genotype H2 (China94-1) at 6.4%-6.9% and from genotype A (Edmonston) at 6.7%-6.9%, from measles vaccine (Shanghail91) at 7.6%-8.0%. They differed from the other measles viral strain isolated in China in 1993 - 2005 at 0.2%-3.7%. The variation within 4 isolated measles viruses was at 0.7%-1.3%.

CONCLUSIONIt was H1 genotype measles viruses,which are the native viruses in China that led to the outbreak of measles in Shanghai, in 2005.

China ; epidemiology ; Disease Outbreaks ; Genotype ; Humans ; Measles ; epidemiology ; genetics ; Measles virus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo determine whether Acanthamoeba polyphaga could affect the survival and growth of Vibrio cholerae O139 in low temperature.

METHODSV. cholerae O139 was co-cultured with the Acanthamoeba polyphaga to be examined on its intracellular growth and survival rate within cysts at low temperature, using methods as Gram-staining, electron microscope and passage culture.

RESULTSV. cholerae O139 were observed to enter into the trophozoites and grow the within the vacuoles after 8 hour incubation with Acanthamoeba polyphaga. The germs survived in the vacuole and/or endo-layer of wall and could be re-isolated from the excystment of Acanthamoeba polyphaga. At 30 degrees C, V. cholerae O139 could survive for 120 days with the amoeba while less than 45 days in PAS. At 4 degrees C, the number of viable bacteria decreased and reached undetectable levels for both study and control groups after a 30-day incubation. V. cholerae O139 could be re-isolated from the 30-, 45-, 60- and 75-day's infected cysts after excystment. However the ability of excystment for 90-day's infected cysts decreased and V. cholerae O139 within the cyst could not be isolated again because the amoebae had lysed.

CONCLUSIONThese findings indicated that V. cholerae O139 could grow within Acanthamoeba polyphaga and the survival time could be increased in the cysts at low temperature. It seemed that Acanthamoeba can provide an environmental reservoir for V. cholerae O139.

Acanthamoeba ; microbiology ; Bacterial Capsules ; Colony Count, Microbial ; Temperature ; Vibrio cholerae ; growth & development

Objective To investigate the effect of high-dose coenzyme Q10 on the central nervous system in mice,and to provide experimental basis for clinical safety evaluation.Methods Mice were randomly divided into vehicle control group,perillartine control group,positive control group (chlorpromazine or diazepam) and coenzyme Q10 low,medium and high dose groups (1.5,3.0 and 6.0 g/kg,equivalent to 75,150,and 300 times of clinical dosage,respectively).The corresponding drugs were ig given to mice with the volume of 40 mL/kg.The general behavior of mice was observed directly,the motor coordination ability was observed by rotating stick method,and Anymaze animal behavior video analysis system was used to observe the spontaneous activity of mice and synergistic reaction with sub-threshold dose of pentobarbital sodium.Results There were no significant differences in the general behavioral activity,and the number of spontaneous activity times,mean resident time,and ratio of sleeping were found in all coenzyme Q10 groups,compared with the vehicle and perillartine control groups.Conclusion High dose of coenzyme Q10 has no significantly toxic effect on the central nervous system in mice,which could provide a reliable experimental basis for further medication study and clinical application of high-dose coenzyme Q 10.

Objective To explore the effects of gravitational traction and target regulation of caspase3 on the degenerative intervertebral disc cells in rabbits.Methods Rabbits nucleus pulposus cells transfected with caspase3 siRNA or negative control siRNA were incubated in serum-starved medium for 6 hours and 48 hours.The expression of caspase3 siRNA and cell apoptosis were analyzed by RT-qPCR and Annexin V-fluorescein staining.In order to create intervertebral disc degeneration model,the right anterior side of the annulus fibrosus of lumbar vertebrae of 35 rabbits were damaged by 16-gauge needle.After confirming the success of modeling,35 animal models were randomized into 3 groups:model group (n=10),negative siRNA group (n=10) and caspase3 siRNA group (n=5).Either negative control siRNA or caspase3 siRNA was injected into the center of nucleus pulposus using a 26-gauge needle from the left anterior side,while the model group received no injection.Nucleus pulposus tissue of 5 rabbits selected randomly from every group after 48 hours were analyzed by PCR and Western blotting for caspase3 mRNA and protein expressions.Half of the casepase3 siRNA group were selected randomly and received a routine gravitational traction using a model of our own design,30min per day for 2 weeks,while other groups received no treatment.TNF-o and IL-1β expression levels and histopathological observations were performed after intervention.Results Expression of caspase3 mRNA in rabbit nucleus pulposus cells transfected with caspase3 siRNA decreased obviously in serum-starved medium,and the apoptosis rate of cells cultured in serum-starved medium decreased significantly (P＜0.05).Caspase3 mRNA and caspase3 protein expression of nucleus pulposus injected by caspase3 siRNA down-regulated in the caspase3 siRNA group.Compared with model group and negative control siRNA group,TNF-α and IL-1β expression levels of caspase3 siRNA traction group decreased significantly (P＜0.05),but there was no statistical significance compared with caspase3 siRNA group (P＞0.05).Pathological observation revealed that viable cell number and extracellular matrix contents increased and collagenous fibers arranged regularly in caspase3 siRNA traction group.Conclusion Gravitational traction and target regulation of caspase3 can prevent apoptotic cell death and delay early intervertebral disc degeneration.

OBJECTIVETo investigate the effect of Chinese medicine (CM) and Western medicine (WM) on quality of life (QOL) after conservative surgery for endometriosis.

METHODSA total of 320 patients with endometriosis were randomized into two groups by using random block design, CM group (160 cases, activating blood circulation and removing blood stasis treatment based on syndrome differentiation) and WM group (160 cases, gonadotropin-releasing hormone agonist or gestrinone treatment) after conservative surgery. Treatment was given for 3-6 months (according to the revised American Fertility Society scoring system stage), and the World Health Organization QOL-BREF (WHOQOL-BREF) was applied to patients before and after treatment to assess QOL.

RESULTSThere were 136 cases in the CM group and 141 cases in the WM group completing therapy. In the CM group, the use of the WHOQOL-BREF showed that the physical, psychological and environmental scores posttreatment were significantly higher than those at pre-treatment (P < 0.05), and for 12 items (pain and discomfort, energy and fatigue, sleep and rest, mobility, activities of daily living, work capacity, negative feelings, health and social care: accessibility and quality, participation in and opportunities for recreation/leisure activities, appetite, QOL score, overall health status and QOL), the difference in scores was significant (P < 0.05). In the WM group, 4 items (pain and discomfort, opportunities for acquiring new information and skills, QOL score, overall health status and QOL) had significantly different scores post-treatment compared with those at pre-treatment (P < 0.05). Before treatment, the QOL in the two groups of patients showed no significant difference (P > 0.05). After treatment, the scores for physical health in the CM group were significantly higher than those of the WM group (P < 0.05) and the scores of 4 items (mobility, activities of daily living, sexual activity, QOL score) in the CM group were significantly higher than those in the WM group (P < 0.05).

CONCLUSIONSCM and WM treatment could improve the QOL of patients with endometriosis after conservative surgery. CM treatment is more effective than WM.

Adolescent ; Adult ; China ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; diagnosis ; surgery ; Female ; Follow-Up Studies ; Gonadotropin-Releasing Hormone ; antagonists & inhibitors ; therapeutic use ; Humans ; Middle Aged ; Pain Measurement ; Pain, Postoperative ; drug therapy ; physiopathology ; Patient Satisfaction ; statistics & numerical data ; Postoperative Care ; methods ; Prospective Studies ; Quality of Life ; Reference Values ; Risk Assessment ; Treatment Outcome ; Young Adult