AIM: To seek a small interfering RNA ( siRNA ) sequence targeting rat inhibitor of nuclear factor kappa Bα ( IκBα) that can specifically and effectively suppress IκBα mRNA expression of rat ciliary muscles in vivo.METHODS:Three IκBα specific double stranded siRNAs were designed and synthesized. They were transfected into rat A7r5 cells which express IκBα gene. Flow cytometry was used to assess transfected efficiency. The mRNA and protein levels of IκBα were examined by Real Time quantitative polymerase chain reaction ( Real Time-PCR ) and western blot to screen a candidate valid sequence with the highest inhibitory rate. The Cy3 labeled non-specific control siRNA or the candidate valid siRNA was then injected into rat anterior chamber. Distribution of Cy3- siRNA in rat ciliary muscles was viewed by fluorescence microscopy, and the inhibitory effect in vivo of the valid siRNA was identified via Real Time-PCR and immunofluorescence. RESULTS: The suppression effect of the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene was most obvious by vitro screening. By anterior chamber injection, this valid siRNA could reach rat ciliary muscles and effectively suppress IκBα gene expression with the highest inhibitory rate of 59. 0% on mRNA level at 24h after RNAi, and 52. 3% on protein level at 72h after RNAi (P<0. 01).CONCLUSION: It proves that the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene is the valid sequence to suppress rat IκBα expression of ciliary muscles by RNAi in vivo.

China ; epidemiology ; Humans ; Incidence ; Pneumoconiosis ; epidemiology

Objective To construct the recombinant eukaryotic expression vector pRNAT-U6,1- siEdg4 which curries small interfering RNA(siRNA)of Edg4 and observe the silencing effect of Edg4 gene targeted siRNA in ovarian cancer cell line SKOV3.Methods The Edg4 gene-targeted hairpin siRNA sequence was designed according to the Edg4 sequence in Genbank,and the two complementary oligo nucleotide strands were synthesized and annealed and inserted into the pRNAT-U6.1 plasmid to build a recombinant Edg4 siRNA eukaryotic expression vector,which was sequenced and identified to contain the correct Edg4 siRNA sequence.The human ovarian carcinoma cell lines SKOV3 were transfeeted with the vector using lipofeetamine method.The efficiency of transfecting cells was observed with fluorescent microscope and the mRNA expression level of Edg4 gene was detected by real time quantitative PCR.The LPA levels in cell supernatants were detected using a biochemical method.And the apoptosis of SKOV3 cells induced by the vector was evaluated by flow cytometry.Results The recombinant eukaryotic expression vector was confirmed to contain correct Edg4 siRNA sequence by PCR and sequencing.After transfection large amounts of green fluorescence were seen in plasma and nuclei of SKOV3 cells and the positive cell rates were 64%.The expression level of Edg4 mRNA in transfeeted SKOV3 cell line was significantly decreased (0.05?0.01 vs 0.29?0.04,P

Objective To establish a quick,economical and reproducible method for high-quality RNA extraction from pancreas.Methods We utilized TRIzol Reagent and liquid nitrogen to isolate total RNA from the rat pancreas.The RNA quality was determined by detection of its content and optic density(A) at 260/280nm,and electrophoresis in 1% non-denatured agarose gel.Then reverse transcription-polymerase chain reaction(RT-PCR) was performed to detect expression of the pancreas-specific genes.Results The content of the total RNA extracted from the rat pancreas reached 3-6?g/mg pancreatic tissues,and A260/280 ratio was 1.75-1.89.Electrophoresis of the total RNA showed 28S and 18S rRNA bands with clear smear between them.The RT-PCR products of pancreas-specific genes including insulin 1,glucagon,?-amylase and housekeeping gene ?-actin all exhibited clear bands on 1% agarose gel,which were located in the expected positions,respectively.Conclusion These results suggest that we have successfully isolated the high-quality and intact RNA from the rat pancreas with TRIzol Reagent and liquid nitrogen.The extracted total RNA can be used in RT-PCR for pancreatic gene expression.

OBJECTIVETo study the antiviral constituents in the stems and leaves of Pithecellibium clypearia.

METHODThe constituents of P. clypearia were systematically separated with various chromatographic techniques in combination with antiviral activity monitoring. Their structures were elucidated by physical and chemical properties and spectral data.

RESULTSix compounds were isolated from P. clypearia and were identified as: tricetiflavan (5, 7, 3', 4', 5'-pentahydroxylflavan) (1), myricitrin (myricetin-3-O-alpha-L-rhamnopyranoside) (2), quercitrin (quercetin-3-O-alpha-L-rhamnopyranoside) (3), quereetin (4), methyl gallate (5) and gallic acid (6).

CONCLUSIONCompound 1 approximately 5 were obtained from this plant for the first time. Compound 4 was found to show an obvious anti-respiratory syncytial virus (RSV) activity.

Antiviral Agents ; chemistry ; isolation & purification ; pharmacology ; Fabaceae ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; Gallic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Inhibitory Concentration 50 ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; chemistry ; isolation & purification ; pharmacology ; Respiratory Syncytial Viruses ; drug effects

Objective To investigate the quality of life of the patients underwent eyeball removal and to analyze their impact factors. Methods The status of 58 patients was investigated with uniform questionnaire.Results The total score of quality of life of 58 patients were (76. 16 ±22.35). Marital status, occupation, economic status is the main factors affecting the quality of life (t =4. 26,3. 86,4. 86;P ＜0. 05). Conclusions We should pay attention to not only the physical treatment, but also the psychological treatment of the patients, to improve the quality of life in patients with eye removal.

Objective To understand psychological experience of eyeball enucleation patients and provide psychological support for and analysis of patients eyeball enucleation psychological experience for clinical nursing staff.Methods Using the form of an interview,interview and analysis data of 24 eyeball enucleation patients.Results Psychological experience of patients with eyeball enucleation mainly include:(1)the desperation and fear;(2)self-image disorder;(3)care and support needs;(4)do righteousness eye demand.Conclusions Patients with eyeball take off have special psychological experience.Nursing staff should pay attention to its inner experiences adopts nursing measures to improve disease cognitive level,reduce the psychological burden and improve its postoperative quality of life.

Objective To establish a performance evaluation index system propitious to nursing management. Methods After reviewing literature, the index system was established and selected based on surrounding theory and expert consulting. Results A set of task pand peripheral performance index were estabilished as the first level index. Including 10 secondary index and 38 third index. Conclusions This new performance appraisal category and system covered the all aspects of influencing nursing quality and can be used to evaluate the performance of nursing job objectively and comprehensively.

Objective To examine the activation and cytotoxieity of human peripheral blood T lymphocyte induced in vitro by human 4-1-BB ligand(4-1-BBL) gene transfected into tumor Tca8113 cells.Methods The eukaryotic expression vector pEGFP-h4-1-BBL was transfected into human oral carcinoma cell line Tca8113 by Lipofectamine<'TM>2000. The transfected cells were then selected in medium containing G-418, cloned by limited dilution and named as Tca8113-4-1-BBL Human 4-1-BBL mRNA and protein expression of transfected cells was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting respectively. The tumor cell vaccines (TCV) were obtained by treatment with mitomycin (MMC). Human peripheral blood mononuclenr cells (PBMC) were prepared from lymphoprep, and then stimulated with anti-CD-3 mAb and incubated with non-transfected or transfected TCV-TcaSI13 cells,respectively. The proliferation of T cells was evaluated by trypan blue exclusion; the CCK-8 was used to detect the cytotoxic effect of T lymphecytes. Meanwhile, the secretion of intefferom-γ (IFN-γ) and interleukin(IL)-2 in culture supernatant was detected by enzyme-linked immunosorbnent assay (ELISA).Results The Tca8113 cells trasfected by pEGFP-h4-1-BBL could express human 4-I-BBL efficiently. As compared with wild type Tca8113 cells, the transfected Tca8113 cells could markedly promote proliferation,IL-2 and IFN-γ production and cytotoxic activity of lymphocytos. Condusions The transfection of human 4-1-BBL gene in Tca8113 cells is effctive in enhancing its immunogenicity and inducing antitumor immune response in vitro.

OBJECTIVETo study the relationship on the prevalence rate of hepatitis B virus (HBV) infection and hepatitis B vaccination in urban citizens aged over 20 years old which would led to the development of strategies on HBV control.

METHODSA total of 3744 subjects from general population were randomly selected in this study. Both ELISA and radio immunoassay were used to test five items of HBV infection, including HBsAg, anti-HBs, HBeAg, anti-HBe and anti-HBc.

RESULTSThe overall standardized infection rate of HBV was 51.7%, and HBsAg, anti-HBs, HBeAg, anti-HBe and anti-HBc were 4.5%, 48.5%, 0.3%, 3.5% and 51.4%, respectively. The two lowest HBsAg positive rates were found in the groups under 30 years old (2.9%) and students (2.6%). Anti-HBc rate among men was significantly higher than seen in women (P < 0.05), and showing a trend of increase with age (chi2 for trend = 256.2, P < 0.001). The standardized rates of HB vaccination in this population was 17.6% and decreasing rapidly with age (P < 0.05). People who had been vaccinated had both lower rates of HBsAg and HBV infection but higher rate of anti-HBs than those who had not (P < 0.05).

CONCLUSIONHB vaccination in adults showed a reducing rate of HBV infection in the general population. Together with the enhancement of expanded program on immunization towards HB vaccination in neonates, much attention should be paid to HB vaccination in adults.

Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Hepatitis B ; epidemiology ; immunology ; prevention & control ; Hepatitis B Vaccines ; immunology ; Hepatitis B virus ; immunology ; Humans ; Immunity, Innate ; Male ; Middle Aged ; Prevalence ; Young Adult