MicroRNAs (miRNAs) are recently discovered major regulators of gene expression, which play a pivotal role in a wide spectrum of biological processes including antiviral defence. There is growing evidence that some viruses either encode their own viral miRNAs or subvert cellular miRNAs. The host-and virus-encoded miRNAs and their targets together thus form a novel regulatory layer of interactions between the host and the virus. A better understanding of host-virus interaction mediated by miRNAs would not only enable us to unravel the molecular basis of viral pathogenesis, but also enable us to develop better therapeutic strategies.

AIMTo prepare rifapentine (RIF) liposomes modified by surfactants for studying their the water-solubility, drug loading effeciency, release rate and pulmonary drug delivery.

METHODSThe film method was used to prepare RIF liposomes. Of verious RIF liposomes morphology by lauric diethanolamide (LDEA), Tween 80 and azone, the properties were studied, envolving morphology, entrapment drug release rate and dissected lung-membrance penetration rate of swine. Pulmonary delivery study was carried out through bronchoscope.

RESULTSThe particle size of RIF-LDEA liposomes was between 15 - 50 nm. The top entrapment efficiency reached 83.0%. The apparent coefficient of membrane penetration (Kp) was 44.29. LD50 was 675 mg x kg(-1) by iv.

CONCLUSIONLDEA increased the water-solubility, loading effeciency and release rate of RIF liposomes. The prepared RIF-LDEA liposomes were suitable for the treatment of pulmonary tubrculosis through bronchoscope.

Animals ; Antibiotics, Antitubercular ; administration & dosage ; chemistry ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; methods ; Ethanolamines ; chemistry ; Lauric Acids ; chemistry ; Liposomes ; Lung ; metabolism ; Particle Size ; Permeability ; Rifampin ; administration & dosage ; analogs & derivatives ; chemistry ; Solubility ; Swine

BACKGROUNDTo repair late median nerve injury, many methods have been used in the past years. The aim of this study was to review a thirteen-year experience in restoration of thumb opposition by transposing flexor pollicis brevis muscle.

METHODSFrom July 1992 to August 2005, 63 patients without thumb opposition because of late median never injury were treated by transposing the flexor pollicis brevis muscle. All the patients had received primary nerve repair after the jnjury. The interval between the injury and the second operation was (1.87 +/- 2.31) years (6 months to 4.2 years). The patients were followed up for 3 to 48 [months mean (22.93 +/- 2.31) months]. A functional evaluation system designed in 1992 were used to estimate the outcomes of the patients.

RESULTSAll the patients gained excellent functional results without complications and disabilities during follow-up.

CONCLUSIONSRestoration of thumb opposition by transposing flexsor pollicis brevis muscle has the following advantages: 1. Operative trauma is minimal; 2. It is not necessary to transpose other tendons; 3. Except for the thumb in opposition, movements of other fingers and the wrist are not restricted postoperatively.

Adolescent ; Adult ; Biomechanical Phenomena ; Female ; Forearm Injuries ; physiopathology ; surgery ; Humans ; Male ; Median Nerve ; injuries ; Middle Aged ; Muscle, Skeletal ; surgery ; Tendon Transfer ; Thumb ; physiopathology ; surgery ; Wrist Injuries ; physiopathology ; surgery

OBJECTIVETo investigate the contribution of latent membrane protein (LMP)1 to nasopharyngeal carcinogenesis via Wnt/β-catenin signal pathway.

METHODSThe recombinant plasmid pHA2-LMP1 was constructed; immunofluorescence staining, Dual-Luciferase Reporter Assay, Western blot and immunohistochemistry staining were used to study the effect of LMP1 on the transcriptional activity and expression of β-catenin.

RESULTS(1) Abnormal expression of β-catenin was obtained in 38 cases (50.7%, 38/75), LMP1 expression was obtained in 38 cases (50.7%, 38/75). There was significantly positive correlation between LMP1 expression and abnormal expression of β-catenin in nasopharyngeal carcinoma tissue (P = 0.008). (2) The expression of β-catenin in nuclei of NPC cell line CNE1 and CNE2 transfected with pHA2-LMP1 plasmid dramatically increased, and the expression was remarkable in poorly-differentiated NPC cell line CNE2 than that of well-differentiated CNE1 cells. (3) LMP1 expression dramatically increased the transcriptional activity of β-catenin in CNE1 and CNE2 cells transfected with pHA2-LMP1 and was in a time-dependent. The transcriptional activity of β-catenin was higher in poorly-defferentiated cell line CNE2 than that of well-differentiated NPC cell line CNE1. (4) LMP1 expression did not affect the total protein expression level of β-catenin in both CNE1 and CNE2 cell lines.

CONCLUSIONEB virus-encoded LMP1 may be involved in the pathogenesis of NPC via β-catenin signal pathway.

Adult ; Aged ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Plasmids ; Recombinant Proteins ; metabolism ; Signal Transduction ; Transcriptional Activation ; Transfection ; Viral Matrix Proteins ; metabolism ; Wnt Proteins ; metabolism ; Young Adult ; beta Catenin ; metabolism

Objective To explore the predictors on the abundance of Rattus (R.) tanezumi in households of commensal rodent plague foci.Methods Thirty natural villages that experienced previous plague cases in Lianghe county,Yunnan province,were selected followed by random selection of 20 households in each village through computer technique.Live traps were set in households to capture small mammals which were then identified to species in the field according to their morphological features.Data on potential factors for abundance of R.tanezumi were collected through questionnaires and field observation and were coded and computerized using EpiData software and further analyzed by hurdle regression model under R software.Results A total of 166 rodents (133 R.tanezumi and 33 Suucus murinus) were captured.Results from final multilevel hurdle regression model showed that the likelihood of R.tanezumi captures increased by 1.67-to 2.76-fold in households belonged to Dai ethnic families that stored foodstuff in metal pails,often raising dogs,and having adjacent houses.The number of R.tanezumi captures increased by 2.18-fold in the villages where over 80％ of the households would raise chickens.In the villages with communal latrine,the likelihood and the number of R.tanezumi capture increased 1.93-fold and 2.38-fold,respectively.While the likelihood of R.tanezumi captures would reduce by 45％-61％ in those households where there were cats and cattle being raised and maize grown in the village.The number of R.tanezumi captures would reduce by 63％ in the households where there were outside toilets.Conclusion The abundance of R.tanczumi seemed to be closely related to the ecological environment factors.Programs on plague control and prevention should relate to ecological factors that influencing the abundance of R.tanezumi.

OBJECTIVETo analyze the susceptibility genes of a Parkinson's Disease (PD) family.

METHODSThe blood samples of a four-generation classic idiopathic PD family were collected. Two-point LOD score method was applied to analyze the linkage disequilibrium between the disease locus and microsatellite markers.

RESULTSWe studied 13 markers near the 9 genes that had been reported to be associated with PD. No obvious evidence showed that the selected markers had anything correlation with PD locus.

CONCLUSIONThese 9 genes are not the susceptibility genes of PD in this family.

Adult ; Aged ; China ; Female ; Genetic Linkage ; Genetic Predisposition to Disease ; Genotype ; Humans ; Microsatellite Repeats ; Middle Aged ; Parkinson Disease ; genetics ; Pedigree

OBJECTIVERecombinant virus pulsated dendritic cells (DCs) may affect their survival, growth and maturity. This study is to test the infection efficiency of recombinant adeno-associated virus carrying hepatitis B core antigen (rAAV-HBV-c) to DCs and the growth and maturity of them.

METHODSPeripheral blood mononuclear cells (PBMCs) were isolated from healthy blood donors. Adherent monocytes were pulsed by rAAV-HBV-c and 293 lysate as controls on the first day of isolation. DCs were cultivated in AIM-V media with 1000 u/ml granulocyte macrophage stimulating factor (GM-CSF), 1000 u/ml interleukin-4 (IL-4) and 50 ng/ml tumor necrosis factor-alpha (TNFalpha) separately in vitro. DCs were examined at different times and the expressions of several clusters of differentiations (HLADR, CD14, CD80, CD83, CD86) were studied using FACS after being cultured for 7 days. The transcription and expression of HBV-C gene were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and intracellular staining fluorescence activated cell sorter (FACS), respectively.

RESULTSThe rAAV-HBV-c infected and uninfected monocytes gradually matured and their morphology had no significant differences. The CDs expressed on the surfaces of the two groups of DCs were also similar (HLADR: 96.1% vs. 94.5%; CD86: 87.7% vs. 89.8%; CD83: 75.6% vs. 78%; CD80: 52% vs. 54.3%; CD14: 6.4% vs. 4.5%). HBV-C gene mRNA expression was measured using RT-PCR and 89.5% of the rAAV-HBV-c infected DCs showed their protein expression using FACS.

CONCLUSIONrAAV-HBV-c can effectively pulse DCs without affecting the growth and maturity of them.

Cells, Cultured ; DNA, Recombinant ; genetics ; Dendritic Cells ; cytology ; immunology ; Dependovirus ; genetics ; Genetic Vectors ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Humans ; Recombination, Genetic

The comparative efficacy of 2 anthelmintics (ivermectin and levamisole) against Baylisascaris transfuga migrating and encapsulated larvae was studied in mice. A total of 60 BALB/c mice inoculated each with about 1,000 embryonated B. transfuga eggs were equally divided into 6 groups (A-F) randomly. Mice of groups A and B were treated with ivermectin and levamisole, respectively, on day 3 post-infection (PI). Mice of groups A-C were killed on day 13 PI. Similarly, groups D and E were treated with ivermectin and levamisole, respectively, on day 14 PI, and all mice of groups D-F were treated on day 24 PI. The groups C and F were controls. Microexamination was conducted to count the larvae recovering from each mouse. The percentages of reduction in the number of migrating larvae recovered from group A (ivermectin) and B (levamisole) were 88.3% and 81.1%, respectively. In addition, the reduction in encapsulated larvae counts achieved by ivermectin (group D) and levamisole (group E) was 75.0% and 49.2%, respectively. The results suggested that, to a certain extent, both anthelmintics appeared to be more effective against migrating larvae than encapsulated larvae. However, in the incipient stage of infection, ivermectin may be more competent than levamisole as a larvicidal drug for B. transfuga.
Animals ; Anthelmintics/*administration & dosage ; Ascaridida Infections/*drug therapy/parasitology ; Ascaridoidea/*drug effects ; Disease Models, Animal ; Female ; Ivermectin/*administration & dosage ; Larva/drug effects ; Levamisole/*administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Rodent Diseases/drug therapy/parasitology ; Treatment Outcome

OBJECTIVETo elucidate the effects of sodium butyrate on rat hepatic oval cell differentiation in vitro.

METHODSHepatic oval cells were isolated from rats fed with a choline-deficient diet supplemented with 0.1% (w/w) ethonine for 4 to 6 weeks. The cultured hepatic oval cells were identified by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). After hepatic oval cells were treated with sodium butyrate, the morphological changes were studied through Giemsa staining and the albumin expression level was tested by Western blot.

RESULTSImmunohistochemical results showed the isolated cells were positive for both mature hepatocyte marker albumin and bile duct cell marker cytokeratin-19. Furthermore, RT-PCR results showed that the cells expressed stem cell marker c-kit, but not hematopoietic stem cell marker CD34. In short, the isolated cells were rat hepatic oval cells. 0.75 mmol/L sodium butyrate induced obvious phenotype changes of hepatic oval cells, including enlargement of the oval cells, a decrease in nucleus to cytoplasm ratio, and a 50% increase in the number of binucleated cells. Western blot results showed that 0.75 mmol/L sodium butyrate markedly raised the expression of albumin.

CONCLUSIONSodium butyrate, a differentiation promoting agent, can induce rat hepatic oval cells (liver progenitor cells) to differentiate into mature hepatocytes in vitro.

Animals ; Butyrates ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hepatocytes ; cytology ; Liver ; cytology ; Rats ; Stem Cells ; cytology

OBJECTIVESElevated tissue inhibitor of metalloproteinase-1 (TIMP-1) expression contributes to excess extracellular matrix in liver fibrosis. This study was designed to construct two recombinant adeno-associated viruses (AAV) carrying antisense RNA and small interfering RNA (siRNA) of TIMP-1 (rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo), and then to compare the suppression of TIMP-1 gene expression on rat hepatic stellate cell (HSC)-T6 cells infected by these two types of viruses in vitro.

METHODSAntisense RNA amplified by rat HSC-T6 and U6 promoter followed by the annealing siRNA were cloned into the AAV vector (pdl6-95/neo) and packed in 293 cells to construct the recombinants rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo. Rat HSC-T6 cells were infected with these recombinant AAVs and selected by using G418, and real-time PCR after reverse transcription and Western blot were performed to detect the transcription and expression level of TIMP-1 gene in these cells.

RESULTSThe results of PCR, restrictive enzyme digestion and gene sequencing confirmed that the pdl6-95/ANTI-TIMP-1/neo and pdl6-95/siRNA-TIMP-1/neo had been reconstructed successfully. After they had been packed in 293 cells to form rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo, they were used to infect HSC-T6. Thirty days after the infection, the transcription level of TIMP-1 in HSC-T6 cells infected by rAAV/siRNA-TIMP-1/neo decreased dramatically compared with the mock control and normal HSC-T6 cells (P less than 0.01), and the expression level of TIMP-1 gene in HSC-T6 cells decreased significantly (60%), while the transcription and expression level of TIMP-1 in HSC-T6 cells infected by rAAV/ANTI-TIMP-1/neo had no significant difference with mock control and normal HSC-T6 cells (P more than 0.05).

CONCLUSIONRNA interference can exert a suppression of TIMP-1 gene in rat HSC, and when this function combines with AAV infection, it can suppress the specific gene expression for a long time by chromosomal integration.

Animals ; Cells, Cultured ; Dependovirus ; genetics ; Genetic Vectors ; Hepatic Stellate Cells ; metabolism ; RNA, Antisense ; genetics ; RNA, Small Interfering ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism