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

This study was conducted to observe bone regeneration guided by poly-DL-latic acid (PDLLA) membrane in rabbit radii bone defects and to explore the mechanism of the membrane guided bone regeneration (MGBR). The animal models of bony and periosteous defects were established in both radii of 40 adult New Zealand white rabbits. The left defect as the experimental side was bridged with PDLLA membrane tube, the right side as the controlled side was untreated. The specimens were collected at 2, 4, 8 and 12 weeks postoperatively. General observation, X-ray, histological observation and biomechanical examination were applied to the repair of the models of MGBR in both groups. Two weeks after operation, with much new bony callus formed outside the tube at both fragments, the membrane tube covered with connective tissues was filled with haematoma and fibrous callus. Twelve weeks after operation, the PDLLA membrane became white and its tube shape was still maintained. However, new bone callus outside the tube almost completely disappeared, and inside the tubes all radii bone defects were successfully repaired with bony union. On the controlled sides, bone defects were filled with connective tissues 2 weeks postoperatively. And 12 weeks after operation, the typical nonunion that had been formed after bone marrow canals were sealed with cortical bone. On the experimental side, the strength of the newly formed bone at the 12th week was higher than that at the 8th week (P<0.05), whereas the biomechanical examination could not be done on the controlled side. Therefore, these findings suggested that the bone regeneration could be successfully guided by PDLLA membrane, and this MGBR technique might be generally used in the treatment of bone defects and nonunion.
Animals ; Biomechanical Phenomena ; Bone Regeneration ; Female ; Guided Tissue Regeneration ; Implants, Experimental ; Lactic Acid ; chemistry ; Male ; Membranes, Artificial ; Osteogenesis ; Polyesters ; Polymers ; chemistry ; Rabbits ; Radius ; injuries ; pathology

Objective: To observe the clinical effect of Tongfu granules(通腑颗粒) on the gastro- intestinal dysfunction in patients with multiple organ dysfunction syndrome(MODS).Methods: The trial was prospective,multi-centric and clinically controlled.One hundred and forty patients with MODS who had been selected were randomly divided into two groups: mosapride citrate group and Tongfu granules group.Respectively at 0 hour,the 48 th hour,the 7 th day or before death, the following scoring systems were calculated: the intestinal dysfunction score,acute physiology and chronic health evaluationⅡ((APACHEⅡ)) score and Marshall score.The duration of mechanical ventilation,hospitalization in intensive care unit(ICU) and the prognosis within 28 days were recorded.Results: After treatments,the intestinal dysfunction score,(APACHEⅡ) score and Marshall score of all patients decreased,at the same time,the therapeutic effects of Tongfu granules group were more significant than those in mosapride citrate group(P20 scores.The mortality was elevated with the increased number of dysfunction organs.Conclusion: Tongfu granules can ameliorate the severity of the disease situation andimprove the prognosis of patients with MODS.

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 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.

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.