Administration, Oral ; Humans ; Rotavirus Infections ; immunology ; prevention & control ; Viral Vaccines ; administration & dosage ; immunology

Adult ; Farmer's Lung ; Female ; Humans

Aim To construct eukaryotic expression vector of shRNA(small hairpin RNA)for human SREBP-1(sterol regulation element binding protein-1)gene and explore its effects on lipid droplet formation in human renal proximal tubular epithelial cell line(HKC)under the stimulation of high glucose.Methods Two eukaryotic expression vectors of shRNA were constructed for human SREBP-1 gene.The HKC cells were transfected with negative control plasmid(pGenesil-1-HK)and two recombinant vectors(pGenesil-1-SREBP1-1 and pGenesil-1-SREBP1-2)and then were cultured under the stimulation of high glucose for about 48 h.The expression of SREBP-1 mRNA and FAS mRNA was detected by RT-PCR and SREBP-1 protein expression was investigated by Western blot.Lipid droplets were detected by Oil Red O staining.Results DNA sequencing showed that the target segments were successfully cloned into pGenesil-1 vector respectively.RT-PCR indicated that two recombinant vectors could inhibit the expression of SREBP-1 mRNA and FAS mRNA in HKC cells under the stimulation of high glucose.Similarly,SREBP-1 protein was also inhibited by the transfection with recombinant vectors.Oil Red O staining found that silencing of SREBP-1 gene resulted in lipid droplets decrease.Conclusions The eukaryotic expression vector of shRNA for human SREBP-1 gene was successfully constructed,and the expression of SREBP-1 was inhibited effectively by the expressed siRNA in HKC cells that resulted in lipid droplets decrease through FAS mRNA transcription inhibition.

Two pairs of primer targeting to the genes encoding the major outer membrane protein (MOMP) and the polymorphic membrane protein (PMP) of Chlamydophila psittaci (Cps) designed and used in the fluorescent quantitative PCR assay in the detection of this microorganism were compared in the their sensitivity and specificity.It was demonstrated that both pairs of primer could be used successfully to detect the presence of Cps of the epidemic strains isolated in China.The specificity of PMP primer used was higher than that of MOMP primer. In case of high target concentration in samples, the sensitivity of the former was also superior to that of the latter, but the amplification efficiency of the former was lower than that of the latter. The MOMP primer seemed to be more suitable to detect Cps by using the real-time quantitative assay.As demonstrated by the real-time quantitative PCR assay, great difference in the genomic copy numbers of the PMP gene existed in the epidemic strains of Cps isolated in China.

The picornavirus family comprises many small viruses, several of which are important pathogens of humans and livestock. The 3C protease (3Cpro) of different species and genera of picornavirus contains the classic G-X-C-G motif and Cys-His-Asp/Glu catalytic triad. 3Cpro conducts maturation cleavage in the regions of VP2-VP3 and VP3-VP1 in P1, 2A-2B and 2B-2C in P2 and the whole P3. Picornavirus 3Cpro has been shown to have significant substrate preference in Q-G/S/A/V/H/R and E-S/G/R/M as well as species and genera specificity through analyses of the maturation cleavage of picornavirus polyproteins. Innate immune adaptors such as TRIF, MAVS, IRF3, IRF7 and NEMO have various potential cleavage sites in picornavirus 3Cpro (TRIF and NEMO show considerable diversity in their cleavage sites). Useful information will be provided for the development of broad-spectrum antiviral agents as well as evasion mechanisms of the innate immune system against picornavirus 3Cpro through continued research of picornavirus 3Cpro.
Cysteine Endopeptidases ; physiology ; Immunity, Innate ; Picornaviridae ; enzymology ; immunology ; Viral Proteins ; physiology ; Virus Replication

Objective To discuss the technical points and clinical effectiveness of the guidewire looping technique for re-canalizing chronic and total long-segment occlusions of femoropopliteal arteries.Methods The guidewire tip was formed into a U-shaped loop and then the guidewire loop was advanced to the occluded artery and was manipulated to re-open the occluded vessel. The catheter followed the guidewire gradually until the tip of the guidewire came into the true lumen of the outflow tract. Results The chronic long-segment occlusions of femoropopliteal arteries were successfully re-canalized in all 48 patients with a technical successful rate of 92.31% (48/51). The re-canalization procedure was failed in three cases.Conclusion Guidewire looping technique is an effective, safe and simple practical skill for re-canalizing chronic long-segment occlusions of femoropopliteal arteries.

ObjectiveTo evaluate the clinical safety and efficacy of SilverHawk directional atherectomy for femoropopliteal occlusive lesions. MethodsEighteen ischemia occlusive lesions in 11 patients of the lower extremity were treated with SilverHawk directional atherectomy.The mean lesion number was 1.6 ± 1. 1 per patient. The mean lesion length was ( 3.4 ± 2. 2 ) cm. The average degree of diameter stenosis was 96％ ± 14％. 9 lesions were totally occlusive. Clinical symptoms included claudication in 4 cases ( Rutherford classes: 3) and critical limb ischemia ( Rutherford classes: 4) in 7 cases. Lesions characteristics were divided by TASC classification: TASC B in 7 cases; TASC C in 1 case (in-stent occlusion); TASC D in 3 cases.Mean ABI was 0. 5± 0.4. Patency was evaluated with color duplex sonography or CTA besides clinical examination during follow-up.ResultsNine totally occlusive lesions were recanalizated successfully via intraluminal approach. 18 lesions achieved technical success (residual stenosis ＜50％ ) leaving 15％ ±7％ mean residual stenosis in mean (8 ±3)min, predilation was needed in one lesion ( in-stent occlusion) prior to atherectomy. Clinical symptoms improved or disappeared with mean ABI 1.07 ±0. 12 and Rutherford grades: 0 (n =9) and 1 (n =2). Patency rate was 100％ with mean 0. 93 ± 0. 14 ABI and Rutherford grades remain unchanged after follow-up of mean ( 9 ± 4 ) monthes.ConclusionsSilverHawk directional atherectomy is safe and effective for the treatment of lower extremity ischemia.

ObjectiveTo evaluate the clinical application of endovascular treatment for severe Takayasu's arteritis (TA).MethodsIn this study,35 target lesions in 32 patients [28 women,mean age (30 ±8) years] with severe Takayasu's arteritis were treated with endovascular merthod.The average length of lesion was 3.1 cm( range 2.7 -5.3).The overall average degree of diameter stenosis was 90％ ± 11％ (range 70- 100)in which 15 lesions were completely occlusive.There were 10 patients whose ESR were higher than 20 mm/h( range 25 -37).Follow-up included physical examination and patency evaluated by color duplex souography/computed tomography angiography/angiography at 6 months and then annually.ResultsRecanalization was unsuccessful in 3 completely occlusive lesions,with a successful rate of 80％(12/15).There was one case in which embolization leading to acute thrombogenesis developed during interventional procedure and resulting in severe stroke.The technical successful rate ( residual stenosis ＜ 50％ ) was 88.6％ ( 31/35 ).The transient cerebral ischemia attack ( TIA ) symptoms disappeared in 31 cases.26 cases were followed up for an average of (19 ± 10) months (range 13 -40).Occipital infarction following severe in-stent restenosis developed 13 months later in one case.Symptomatic in-stent restenosis18monthslaterwasfoundin2cases. Patencyratewas88.5％( 23/26 ).ConclusionsEndovascular treatment is safe and effective for severe TA.Strict indication and accurate targeting the lesions help ensure the success of management.

To developing a simple, rapid and sensitive immunoaffinity method for extracting DNA before amplification by the polymerase chain reaction(PCR). Using monoclonal anti-DNA bound to colloidal-gold beads(which called immunocolloidal-gold ) and extracting DNA from the specimen before amplification by PCR. The immunoaffinity method we use is extremely efficient in extracting DNA at low concentration, also, the beads carrying the DNA/anti-DNA complexes can be added directly to a PCR mixture without elution, and the presence of PCR inhibitors in specimens has no effect on amplification. When applied to serial dilution bacteria suspension, the detection of limit can be 100 CFU/mL, and the sensitivity of detecting artificial soil sample and milk sample were 101 and 100 CFU/mL. The immunoaffinity method described here can sever as a sensitive and practicable method for extracting DNA , particularly where samples have low concentrations of DNA or where the material is in poor condition.

A CdTe/ZnSe quantum-dot submicrobead ( QBs ) , which exhibited fluorescence intensity approximately 2800-fold stronger than that of single quantum dots, was conjugated with the anti-histidine rich protein( HRP )-Ⅱ mAbs using N-( 3-( Dimethylamino ) propyl )-N'-ethylcarbodiimide hydrochloride ( EDC ) method as fluorescence probe. The goat anti-HRP-Ⅱ polyclonal antibodies and donkey anti-mouse polyclonal antibodies were sprayed onto the nitrocellulose membrane as test line and control line, respectively. The resultant fluorescence probes were introduced to the immunochromatographic strip for the quantitative determination of Plasmodium falciparum. For determination of Plasmodium falciparum in serum, the QBs based immunochromatographic strips exhibited a good dynamic linear range from 5 . 8 Parasite/μL to 8010 Parasite/μL with a limit of detection of 5. 8 Parasite/μL. The detection time of the proposed QBs based immunochromatographic strips for each sample was only 15 min. Moreover, the recovery rates of the intra-and inter-assay ranged from 93. 0% to 111. 9%, and 98. 3% to 115. 1% respectively, while the relative standard deviations ( RSDs) of intra-and inter-assay were below 5%.