Animals ; Brain ; drug effects ; metabolism ; Carbon Dioxide ; metabolism ; Chronic Disease ; Curcumin ; pharmacology ; Disease Models, Animal ; Fas Ligand Protein ; metabolism ; Hypoxia ; metabolism ; Male ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; fas Receptor ; metabolism

Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions. The GeXP system after optimization could amplify the specific fragments related to the viruses and exposed specifically a total of 13 target genes out of eight types of arboviruses at the level of 10(2) copies/microL, and the findings suggest that the novel protocol we developed can be high-throughput and highly specific and sensitive as well as quickness in screening of the encephalitis viruses, and is promising in detection of encephalitis-associated viruses for molecular epidemiological studies.
Arboviruses ; genetics ; isolation & purification ; Encephalitis Virus, Japanese ; genetics ; Encephalitis Viruses ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To evaluate the effect of up-converting phosphor technology(UPT) in detection of plague antigen-antibody by receiver operating characteristic curve (ROC) method,and to provide a scientific basis for field application of UPT rapid detection technology in plague prevention and control.Methods Two hundred and twenty four serum samples were collected from Marmots and ground squirrels in the plague foci,Yersinia pestis antibody was detected by UPT,ELISA,Colloidal-gold Strips and IHA,respectively; 108 organs and bone marrow samples were collected,and Yersinia pestis antigens were detected by UPT,ELISA,PCR and RIHA,respectively.IHA was used as the gold standard for antibody test results,RIHA,PCR + Colloidal-gold Strips,PCR + ELISA were used as the gold standard for antigen test results.The results were evaluated using ROC method.Results Antibodies detection:the AUCs of UPT,ELISA and Colloidal-gold Strips were greater than 0.5.The difference between UPT and other methods was not statistically significant (z =1.204,P ＞ 0.05).Antigen detection:the AUCs of UPT,ELISA,Colloidal-gold Strips and PCR were greater than 0.5.There was no statistical difference between UPT and other methods(z =0.866,P ＞ 0.05).Conclusions UPT as a new technology works well in the detection of plague antigen-antibody.The technology is simple,fast,accurate,and suitable for on-site monitoring of plague,emergency treatment of sudden plague,and suitable for promotion.

OBJECTIVETo reveal the genetic characteristics of GII.12 Norovirus strains isolating from stool samples of adults with diarrhea in Beijing during 2008-2009.

METHODSRdRp, ORF2, ORF3 and ORF1/ORF2 overlap region were respectively amplified by primers using RT-PCR. The products were purified, cloned, sequenced and then aligned, phylogenetic and recombinant analyzed by softwares of DNAStar, MEGA and SimPlot.

RESULTSAccording to the phylogenetic analysis, 11 strains belonged to G II.g in the RdRp region,while GII.12 in the ORF2 and ORF3. SimPlot analysis further confirmed the 11 strains were recombinant strains ( G II.g [RdRp]/G II.12 [capsid]).

CONCLUSIONG II.12 Norovirus prevailing in Beijing and other regions of the world belonged to the same strain, and we identified the genetic characteristics of G II.12 Norovirus in Beijing.

China ; Norovirus ; classification ; genetics ; Phylogeny ; Recombination, Genetic ; Time Factors

OBJECTIVETo obtain information on viral molecular structural and evolutionary characteristics, we conducted the SZ2010422 full-length genomic analysis.

METHODSPrimers were designed by New Orleans full sequence, SZ2010422 full genome was amplified by RT-PCR, the whole genome sequence and the capsid domain amino acid sites was analysised after cloned and sequenced.

RESULTSThe genome of G II-4 Norovirus SZ2010422 strain was consist of 7559 bp, it revealed three ORFs composites of the whole genome, ORF1 (5100 bp), ORF2 (1623 bp), ORF3 (807 bp) respectively, ORF1 and ORF2 had 19 nucleotide overlap. By evolutionary comparative analysis found SZ2010422 genomic nucleotide sequences with reference strains of G II-4 New Orleans1805 strains the highest homology with a total length of homology was 99.3%, of ORF1 (99.5%), ORF2 (99.2%), ORF3 (98.6%). Phylogenetic analyses showed SZ2010422 belonging to G II-4 New Orleans variant. Date of 541 amino acid analyses showed: New Orleans variant strains of popular sites: aa310N or K, --> S aa341D --> of N, aa359T--> S, aa396H --> P, aa460H --> Y.

CONCLUSIONNorovirus SZ2010422 belonged to the G II-4 New Orleans variant. In This study, SZ2010422 full sequence can be used not only as a full-length NoV variant sequence standard for future comparison studies, but also as useful material for the public health field by enabling the diagnosis, vaccine development, and prediction of new emerging variants. Noroviruses; Genes; Sequence analysis

China ; Genome, Viral ; Norovirus ; classification ; genetics ; Open Reading Frames ; Phylogeny ; Sequence Analysis, DNA

BACKGROUNDThe evidence for non-invasive positive pressure ventilation (NIPPV) used in patients with severe stable chronic obstructive pulmonary disease (COPD) is insufficient. The aim of the meta-analysis was to assess the treatment effects of long-term NIPPV on gas change, lung function, health-related quality of life (HRQL), survival and mortality in severe stable COPD patients.

METHODSRandomized controlled trials (RCTs) and crossover studies comparing the treatment effects of NIPPV with conventional therapy were identified from electronic databases and reference lists from January 1995 to August 2010. Two reviewers independently assessed study quality. Data were combined using Review Manager 5.0. Both pooled effects and 95% confidence intervals were calculated.

RESULTSFive RCTs and one randomized crossover study with a total of 383 severe stable COPD patients were included. NIPPV improved gas change significantly when using a higher inspiratory positive airway pressures. The weighted mean difference (WMD) for the partial pressure of carbon dioxide in artery (PaCO2) was -3.52 (-5.26, -1.77) mmHg and for the partial pressure of oxygen in artery (PaO2) 2.84 (0.23, 5.44) mmHg. There were significant improvements in dyspnea and sleep quality, but gained no benefits on lung function. The standardized mean difference (SMD) for the forced expiratory volume in 1 second (FEV(1)) was 0.00 (0.29, 0.29). And the benefits for exercise tolerance, mood, survival and mortality remained unclear.

CONCLUSIONSPatients with severe stable COPD can gain some substantial treatment benefits when using NIPPV, especially improvements in gas change, dyspnea and sleep quality. Studies of high methodological quality with large population, especially those based on a higher inspiratory positive airway pressures are required to provide more evidences.

Aged ; Humans ; Positive-Pressure Respiration ; Pulmonary Disease, Chronic Obstructive ; therapy

OBJECTIVETo investigate the effects of drug-containing serum of Chinese herbal compound, Xiongshao Capsule (, XS, for activating-blood) and Huanglian Capsule (, HL, for dispellingtoxin) on the oxidized low-density lipoprotein (ox-LDL)-induced inflammatory factors in human umbilical vein endothelial cells (HUVECs).

METHODSThirty-two rats were randomly divided into four groups: the blank control group treated with distilled water, the positive control group treated with simvastatin (1.8 mg/kg), the test group I treated with Chinese herbal compound of XS (0.135 g/kg), and the test group II treated with Chinese herbal compound of XS (0.135 g/kg) and HL (0.135 g/kg). All the treatments were administered for 7 successive days by gastrogavage. Rats' blood serum was harvested 1 h after the last administration to prepare respective drugcontaining serum. HUVECs were exposed to ox-LDL (100 μg/mL) to induce cell injury model and incubated with corresponding drug-containing serum for 24 h. Untreated HUVECs were set for blank control. Levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and soluble intercellular adhesion molecule-1 (sICAM-1) in supernatant of cultured HUVECs were determined by enzyme-linked immunosorbent assay (ELISA). HUVEC surface expressions of ICAM-1 and E-selectin were determined by flow cytometry.

RESULTSLevels of IL-6, TNF-α, and sICAM-1 in the supernatant of HUVECs as well as the cell surface expressions of ICAM-1 and E-selectin significantly increased after 24-h ox-LDL stimulation (P<0.01), while the abnormal elevations, except sICAM-1 in the test group I, were all reduced in the treated groups (the positive control and the two test groups) significantly (P<0.01 or P<0.05). Besides, the effect in the test group II seemed somewhat higher than that in the test group I but with no statistical significance (P>0.05).

CONCLUSIONDrug-containing serum of XS plus HL has a certain inhibitory effect on the vascular endothelial inflammation response induced by ox-LDL.

Animals ; Capsules ; Cell Membrane ; drug effects ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; E-Selectin ; metabolism ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Inflammation Mediators ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-6 ; metabolism ; Lipoproteins, LDL ; metabolism ; Rats ; Rats, Wistar ; Solubility ; drug effects ; Subcellular Fractions ; drug effects ; metabolism ; Toxins, Biological ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo construct eukaryotic expression vectors by using the pSilencer3. 1-H1 neo vector for inhibiting human survivin gene by RNA interference, and to detect the effect of the silenced survivin gene on PC-3 cells.

METHODSThree target gene segments were synthesized and cloned into the pSilencer3. 1-H1 neo vector respectively to construct three recombinant eukaryotic expression vectors: pSilencer3. 1-SVV1, pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3, which were identified by enzyme digestion analysis and DNA sequencing. Then the PC-3 cells were transfected with the recombinant vectors and the interference effect detected by RT-PCR, Western blot and immunohistochemical staining. The apoptosis index of the PC-3 cells was detected by flow cytometry and their proliferation detected by MTT method.

RESULTSEnzyme digestion analysis and DNA sequencing showed that three target segments were cloned into pSilencer3. 1-H1-neo vectors. The results of RT-PCR, Western blot and immunohistochemical staining indicated that pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3 vectors could knock down the transcription and expression of survivin gene. After transfected with pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3 vectors, the apoptosis index of the PC-3 cells was increased by 10% - 15% and their growth obviously slowly down.

CONCLUSIONThe transcription and expression of survivin gene were inhibited effectively by the recombinant eukaryotic expression vectors (pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3) in the prostate cancer cell line PC-3. After transfected with pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3 vectors, the apoptosis index of the PC-3 cells was increased and their growth inhibited.

Apoptosis ; Cell Line, Tumor ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; drug effects ; Genetic Vectors ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Prostatic Neoplasms ; metabolism ; pathology ; RNA Interference ; RNA, Small Interfering ; pharmacology ; Transfection

A randomized crossover study was performed to compare the effects of low glycemic index diets (LGI)and high glycemic index diets(HGI)on blood glucose,lipid profile and control of body weight in patients with type 2 diabetes.Compared with HGI group,the fasting serum insulin,Homa-IR,LDL-C and body weight significantly decreased in LGI group(P

[ Objective ] To observe the effect of kaolin and propranolol on paraquat ( PQ ) concentration in the lungs of poisoned mice. [ Methods] A group of 144 ICR mice were randomly divided into 3 groups:PQ, treatment, and control.Then 100 mg/kg PQ were intragastrically administrated ( ig) in PQ group and treatment group, while only the same volume normal saline was given in control group.And then 48 g/kg kaolin combined with 3.2 mg/kg propranolol were administered in treatment group immediately after poisoning while only the same volume of normal saline was given in the other two groups. Pathological examination was done and PQ concentration in lungs of the mice detected 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h after poisoning. [ Results] In the lungs of the mice in PQ group occurred alveolar capillary expansion, endothelial cell swelling, small or large sheet-shaped inflammation cell infiltration and mainly neutrophils while in treatment group the above lesions were apparently alleviated.In PQ and treatment groups, PQ concentration in lungs of both groups rose significantly 0.5 h after poisoning and up to peak at 4 h.But PQ concentration in lungs decreased significantly in treatment group from 4 h to 24 h after poisoning ( P<0.05) , as compared with that in PQ group. [ Conclusion] PQ concentration in lungs of the poisoned mice was decreased and the injury alleviated when they were treated with kaolin combined with propranolol.It is held that further research is worth doing in clinical practice.