ObjectiveTo compare the therapeutic effec between sustained low-efficiency dialysis (SLED) and continuous blood purification (CBP) in critically ill patients.MethodsAccording to the treatment ways,96 critically ill patients were divided into SLED group and CBP group.A comparison was made on the biochemical indicators,in-hospital duration,hemodynamic parameters,acute physiology and chronic health evaluation (APACHE-Ⅱ ),the survival and the mortality rates.ResultsAfter treatment,the levels of serum creatine kinase isozyme MB (CK-MB),creatine kinase (CK),creatinine (Cr),glutamic-oxalacetic transaminase (AST),glutamate-pyruvate transaminase (ALT),APACHE Ⅱ score on the 1st,2nd and 7th day were lower than those prior to the treatment in both groups ( P ＜0.05).There were no statistical differences in in-hospital duration, biochemical indicators, APACHEⅡscore,hemodynamic parameters,the survival rate and the mortality rate between the two groups (P ＞ 0.05 ).ConclusionsSLED has similar hemodynamic stability with CBP,and the two methods have similar treatment effects in critically ill patients.However,SLED can be relatively economical and convenient for critically ill patients in clinical.

RNA interference (RNAi) is a promising technology in development of specific antiviral therapy, but the quantitative detection of small interfering RNA (siRNA) expressed in vivo is the main challenge to assess its antiviral effect. In order to detect the siRNA molecules (siN1 and SiN2) particularly expressed in cells to inhibit the replication of classical swine fever virus (CSFV), serial specific stem-loop primers were designed and synthesized. Two of them (SLP-N1-6 and SLP-N2-8) were selected by screening in cross combination and successfully used in establishment of an optimal stem-loop RT-qPCR, which showed high specificity and sensitivity in detection of anti-CSFV siRNA expressed in PK-15 cells. The method was capable of detecting 10(2) to 10(8) copies of siRNA molecule with good parallel relationship (R(sq) = 0.999) and high amplification efficiency (Eff. = 98.2%). Therefore, the established stem-loop RT-qPCR can be used as an ideal tool in quantitative assessment of the anti-CSFV effects of RNAi in combination with detection of viral antigens using indirect immunofluorescent assay and TCID50, providing a novel technique for evaluating the antiviral effects of the siRNA expressed in anti-CSFV transgenic pigs to be established in future.
Animals ; Cell Line ; Classical swine fever virus ; genetics ; isolation & purification ; metabolism ; RNA Interference ; RNA, Small Interfering ; analysis ; genetics ; metabolism ; RNA, Viral ; genetics ; Real-Time Polymerase Chain Reaction ; methods ; Swine ; Transfection ; Viral Nonstructural Proteins ; genetics ; metabolism ; Virus Replication

OBJECTIVETo explore the expression of hsa-mir-186 in colorectal cancer and study its role in regulating the biological behaviors of human colorectal cancer SW620 cells in vitro.

METHODSThe expression of hsa-miR-186 in colon cancer tissue and the adjacent tissues as well as 5 colon carcinoma cells were analyzed using real-time quantitative RT-PCR. The precursor sequence of miR-186 gene was amplified from the genomic DNA by PCR and cloned into the lentiviral vector PLVTHM labeled with GFP. The colorectal cancer cell line SW620 was transfected with PLVTHM-miR186 vector and the lentivirus-infected cells were sorted with flow cytometry. Cell counting kit-8 (CCK-8) assay was used to detect the proliferation of the cells. The migration and invasion of SW620 cells were investigated using Transwell assay and scratch test. Western blotting was used to detect the expression of YY1 protein in SW620 cell lines.

RESULTSThe relative expression of miR-186 in the cancer tissues was 0.0024∓0.0027, significantly lower than that in the adjacent tissues (0.066∓0.068, P=0.008); the relative expression level of hsa-miR-186 in SW620 and LoVo cells with a high metastatic potential was 0.118∓0.138 and 0.157∓0.001, respectively, significantly lower than that in HT-29 cells with a low metastatic potential (1.000∓0.00, P<0.05). The recombinant lentiviral vector PLVTHM-miR186, verified by enzyme digestion, sequencing and qPCR, caused significant inhibition of cell proliferation, migration and invasion and suppressed the expression of YY1 protein in SW620 cells.

CONCLUSIONAs a tumor suppressor gene, Hsa-miR-186 is down-regulated in colon carcinoma tissues and in highly metastatic SW620 and LoVo cells. Has-miR-186 can inhibit the cell proliferation, migration and invasion of colon carcinoma cells in vitro possibly by suppressing YY1 expression.

Adult ; Aged ; Aged, 80 and over ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colonic Neoplasms ; genetics ; pathology ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; MicroRNAs ; Middle Aged ; Transfection ; YY1 Transcription Factor ; metabolism