2.pRNA: A Safe,Stable and Large Capacity Molecular Vehicle for Gene Therapy
Feng-Ping XU ; Jun YANG ; Xue-Song FENG ; Li-Jia AN ;
China Biotechnology 2006;0(06):-
The molecular delivery vectors used in gene therapy need provide the features of safety,stability, efficiency and capacity. The current studies on the structure and action mechanism of pRNA, a packaging RNA of phage?29, showed that pRNA with multiple binding sites can through cell membrane easily and escort exogenous molecules to target cell, without inducing immune reaction. As an ideal nano-scale gene therapy vehicle, pRNA presents a promising application in delivering multiple therapeutic components to detect and treat human diseases.
3.Investigation on the influence of the ROX reference fluorescence correction on the real-time quantitative RT-PCR assessment for Nrf2 mRNA
Feng GAO ; Xuewen YANG ; Jia WANG ; Chunbing ZHANG
Chinese Journal of Laboratory Medicine 2011;34(8):742-747
Objective To evaluate the normalized correction effect of the ROX reference fluorescence on the real-time quantitative RT-PCR assessment for the Nrf2 mRNA.Methods The Taq-man probe was utilized in the real-time quantitative RT-PCR method to assess the Nrf2 mRNA; The reaction specificity of the Nrf2 PCR products obtained from the rat cDNA was assessed using gel electrophoresis and sequencing identification.The PCR calibrators with different concentrations varied at 10 gradients(from 2.0 × 109 to 2.0 × 100) were prepared for real-time quantitative RT-PCR assessment,and then the influence of the ROX reference fluorescence correction on the linear range of the PCR standard curve was investigated experimentally.The recombinant plasmids with different concentration levels(e.g,high,medium or low concentration) were assessed by the real-time RT-PCR and 20 replicate measurements were performed for all the samples in both intra- and inter-assays.The repeatability differences of both the inter- and intra-assays were analyzed systemically with/without the ROX reference fluorescence correction.Results The primers and probes required for Nrf2 mRNA real-time quantitative RT-PCR assessments were constructed,and the reaction system and conditions were also established.The length of the amplified fragment agreed well with the expected length(122 bp),and the sequencing analysis showed the amplified fragment was verified by the sequencing identification.After applying the ROX reference fluorescence correction,the linear range ofthe PCR standard curve was measured as 2.0 × 109 -2.0 × 100 copies/μl(R2 > 0.99),which was 100times wider than that measured without ROX reference fluorescence correction(2.0 × 109 - 2.0 × 102copies/μl,R2 > 0.99).Without the ROX reference fluorescence correction,the intra-assay CVs of the Ct value were measured as 4.1%,2.7% and 2.1% for the recombinant plasmids with high-,medium-,and low-level concentration,respectively.The inter-assay CVs of the Ct value were 4.3% 、3.0% 、2.4%.By contrast,with the ROX reference fluorescence correction,the intra-assay CVs of the Ct value became 0.7%,0.5%,0.4% and the inter-assay CVs of the Ct value turned to 1.0%,0.8% and 0.7%.The discrete degree of the Ct value with the correction was lower than that without the correction.The intra- and interassay Ct values of the high- and medium-level samples exhibited statistic significance for the measurements with/without the correction.In the high-level group,the t values of the intra- and inter-assays were 4.843 and 2.566,with P<0.05.In the medium-level group,the t values of the intra- and inter-assays were 4.293 and 4.423,with P<0.05.However,no statistic significance was observed the low-level group(the t values of the intra- and inter-assays were 0.753 and 1.279,with P >.0.05).Conclusions The utilization of the ROX reference fluorescence in the real-time quantitative PCR assessment for Nrf2 mRNA could widen the linear range of the standard PCR calibration curve and improve the repeatability of the assessment,which should be quite helpful for accurately assessing the expression level of the Nrf2 mRNA.
4.Effect of gap junction on permeability of blood-brain barrier in rats after cerebral ischemia- reperfusion
Dong HAN ; Jia-Chun FENG ; Fang DENG ; Yi-Min YANG ;
Chinese Journal of Neurology 2005;0(07):-
Objective To investigate the possible mechanism of the gap junctional influence on the change in permeability of the blood-brain barrier(BBB)after reperfusion subsequent to cerebral ischemia.Methods In the test laser scanning confocal microscope(LSCM)was used to investigate the change of Cx43 levels and distribution.The MCAO/R model was induced using intraluminal suture technique first described by Longa with a little modification.A total of 60 Wistar rats were divided into 4 groups:the sham-operation group,control group,octanol-treatment group and DMSO vehicle control group. Control group were further divided into seven subgroups at different time points of reperfusion after middle cerebral artery occlusion.To observe the change in permeability of BBB,Evans blue(EB)in the brain tissue was surveyed by the means of EB fluorescent quantitation.Octanol-treatment group and DMSO vehicle control group were done at the point of the peak of permeability of BBB.Octanol,the specific blocker for gap junctions(GJ)was used in an intervention study.To compare the amount of EB with the same point of groups,the influence of octanol on BBB permeability was investigated.Results At 3 h of reperfusion after cerebral ischemia for 2 h,the permeability of BBB began to increase,reached the peak at 24 h of reperfusion and was still elevated at 72 h.The Cx43 expression formed into bigger plague and remained linear disposition in the penumbra after reperfusion subsequent to cerebral ischemia.Octanol group was done at 24 h of reperfusion after cerebral ischemia.The amount of EB of octanol group((4.924?0.296)?g/g)was significantly lower than that of corresponding operation control group(5.543?0.506)?g/g.Conclusions (1)Cx43 expression is concentrated around vessels in brain.The Cx43 forms into bigger plague and the function maybe strengthens after reperfusion.Gap junction might aggravate the disruption of BBB.(2) Octanol,the specific blocker of gap junctions,could effectively prevent the permeability of BBB from increasing and has a protective effect on BBB.
5.Novel no-flip Shang Ring circumcision for adult males: a clinical application study of 528 cases.
Ben-Hai YANG ; Chao JIA ; Tao LIU ; Yi-Feng PENG
National Journal of Andrology 2014;20(8):709-714
OBJECTIVETo evaluate the safety and clinical outcomes of the novel simplified no-flip surgical approach to Shang Ring male circumcision in adults.
METHODSA total of 528 adult males, aged 18 - 58 (mean 35) years, 63 with phimosis and 465 with redundant prepuce, underwent no-flip Shang Ring circumcision with or without removal of the outer ring. The operation time and intraoperative blood loss were recorded and observations were made on such complications as postoperative bleeding, infection, edema, and wound dehiscence.
RESULTSThe operation time, intraoperative blood loss, and 2-hour postoperative pain score were (3.8 +/- 0.3) min, (0.6 +/- 0.1) ml, and 7.3 +/-0.3, respectively. Spontaneous ring detachment occurred at 21.6 +/- 2.1 days postoperatively in 12.7% of the patients (67/528) who had chosen not to remove the rings. Of those who preferred removal of the outer ring (87.3% [461/528]), none experienced any pain at the ring removal 7 days after the operation. Postoperative complications included infection in 3 cases (0.56%) and mild edema in 9 (1.70%), but no bleeding and wound dehiscence. Totally, 518 (98.1%) of the patients felt satisfied with the postoperative penile appearance.
CONCLUSIONNo-flip Shang Ring circumcision, with no need for removal of the inner ring, is a safe and simple approach for adult males, which is superior to conventional Shang Ring circumcision for requirement of fewer surgical instruments, shorter operation time, lower incidence of complications, and better satisfaction with the penile appearance. However, these advantages are to be further demonstrated by more randomized controlled trials.
Adolescent ; Adult ; Circumcision, Male ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Phimosis ; surgery ; Postoperative Complications ; Young Adult
6.Clinical Analysis of Thyroid Carcinoma(report of 146 casees).
Haidong CUI ; Zhong JIA ; Yang CAI ; Zhongyao LUO ; Guanghua FENG
Journal of Medical Research 2006;0(01):-
Objective To analyze incidence of thyroid carcinoma and to explore and conclude the principle of its diagnosis and treatment.Methods One hundred and forty six patients of thyroid carcinoma in our hospital were analyzed and discussed with literature.Results 101 cases were diagnosed as thyroid carcinoma by pathological diagnosis of fast frozen sections,the positive ratio is approximately 93.5%.totle resection of affected lobe and isthmus was performed on 35 cases,total resection of affected lobe and isthmus and majority of opposite lobe was done on 37 cases.The lymph node metastasis was found by postoperative pathological diagnosis with a rate of 32.5%.Conclusions Preoperative diagnose of thyroid carcinoma is rarely available,diagnosis of thyroid carcinoma by fast frozen sections is optimal.total resection of affected lobe and isthmus or total resection of affected lobe and isthmus and majority of opposite lobe are the main operation patterns.
7.Compare of Selectivity Enrichment Broth for Detectable Effect of Listeria monocytogenes
Yang-Feng HU ; Jun HAN ; Ying-Min JIA ;
Microbiology 2008;0(12):-
This paper investigated contamination situation of Listeria monocytogenes(Lm). To compare dif- ferent selectivity enrichment broth for detectable effect of Lm and compare detectable effect in different samples by using different methods, furthermore, choose the best enrichment broth for specific food. One hundred and thirty five random samples from raw meat, aquatic product, fruit and vegetable, quick-frozen food in Baoding. Applied LB enrichment broth, EB enrichment broth, new modification FDA enrichment broth and Fraser enrichment broth before separated by PALCAM selective agar, then identified by interna- tional standard method after PCR. Results: Four methods showed that there were 23 Lm positive, detected 5 Lm by LB method, 6 Lm by Fraser method, 5 Lm by EB method and 7 Lm by new modification FDA method. The total detectable rate of four methods had no large specificity, but to specific kind of food was different.
8.Diffusion-weighted MRI of the breast:lesion characterization and parameter selection
Ya-Jia GU ; Xiao-Yuan FENG ; Feng TANG ; Wei-Jun PENG ; Jian MAO ; Wen-Tao YANG ;
Chinese Journal of Radiology 2001;0(05):-
Objective To evaluate the b value of diffusion-weighted(DW)MRI in distinguishing between benign and malignant breast lesions.Methods Three diffusion-weighted sequences were implemented with 500,1000 and 2000 s/mm~2 b values respectively on 95 breast lesions in 83 patients.All lesions were confirmed by pathology.The apparent diffusion coefficient(ADC)values and signal intensity (SI)were recorded and compared in different lesions(breast cancer,benign lesion,cyst and normal beast tissue)with the same b value and the same lesions with the different b values.Results(1)The mean ADC value and SI of breast cancer were 1.375?0.378 and 839.713?360.493 respectively with b= 500 s/mm~2,1.176?0.311 and 459.314?229.609 with b=1000 s/mm~2,0.824?0.198 and 243.825? 110.616 with b=2000 s/mm~2.The differences in the mean ADC value were significant between two type lesions(cancer and benign lesion,cancer and cyst,cancer and normal breast tissue)with b values of 500 s/mm~2 and 1000 s/mm~2.But the significant differenee was only seen between cancer and benign lesions when b value was 2000 s/mm~2.(2)The one-side upper limits of 95% confidence interval of mean ADCs were adopted as the point to separate the malignant from the benign lesions,the sensitivity was 70.92%, 70.73% and 69.77%,the specificity was 77.19%,75.70% and 54.76%,the accuracy was 77.12%, 74.32% and 62.35% respectively with b values of 500 s/mm~2,1000 s/mm~2 and 2000 s/mm~2.The areas under ROC eurves were Az_(500)=0.775?0.046(P0.05).Conclusion DWI MRI is useful for the differential diagnosis of breast lesions with b values of 500 s/mm~2 and 1000 s/mm~2.
9.Analysis of immunophenotypic features of blasts in patients with myelodysplastic syndrome by flow cytometry and its diagnostic significance.
Jia-Feng SUN ; Bo YANG ; Li ZHANG ; Yan HUANG ; Jia YANG
Journal of Experimental Hematology 2012;20(3):632-635
This study was purposed to analyse the characteristics of blast immunophenotype in patients with refractory anemia with excess blasts, type 1 (RAEB-1) and refractory anemia with excess blasts, type 2 (RAEB-2) by flow cytometry and investigate its diagnostic significance, as well as to compare the blast rate detected by FCM and bone marrow smear (BMS). FCM was used to count the blast rate and detect the expression of its antigens in 29 cases of MDS. The result indicated that: (1) The rate of the blasts detected by FCM and BMS was not statistically significant between patients with RAEB-1 and with RAEB-2 (P > 0.05); (2) Out of 13 patients with RAEB-1, the blasts of 10 cases, 12 cases, 8 cases, 11 cases, 11 cases, 3 cases, 7 cases, 0 case, 0 case, 3 cases and 2 cases patients had positive expressions of CD34, HLA-DR, CD117, CD33, CD13, CD15, CD11b, CD3, CD19, CD7 and CD56, separately. The blasts of 12 cases, 13 cases, 3 cases, 12 cases, 15 cases, 7 cases, 5 cases, 0 case, 1 case, 4 cases and 2 cases among 16 patients with RAEB-2 positively expressed CD34, HLA-DR, CD117, CD33, CD13, CD15, CD11b, CD3, CD19, CD7 and CD56, respectively. However, there was no significant difference in the expression of antigens in blasts of the bone marrow between the patients with RAEB-1 and with RAEB-2 (P > 0.05); (3) The positive expression for CD15 and CD11b in blasts was found in 10 cases and 12 cases, respectively; (4) The positive expression for CD19, CD7 and CD56 was observed in 1, 7 and 4 cases, respectively. None of the 29 cases of MDS was positive for CD3. It is concluded that FCM can reveal the characteristics of immunophenotypic abnormalities of the blasts in patients with MDS and provide the important information for diagnosis and differential diagnosis of MDS.
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Adult
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Aged, 80 and over
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Bone Marrow
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pathology
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Flow Cytometry
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Immunophenotyping
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Myelodysplastic Syndromes
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pathology
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Young Adult
10.Study on expression of long non-coding RNA in rheumatoid arthritis
Yan XIA ; Jia FENG ; Anping CHEN ; Nianan YANG ; Yang XIANG ; Linchong SU ; Lin YUAN
Chinese Journal of Immunology 2016;(1):9-12,18
Objective:To investigate the expression profile variation of long non-coding RNA( lncRNA) in the peripheral blood mononuclear cells of rheumatoid arthritis ( RA ) and healthy controls, and explore the role of lncRNA in the pathogenesis of RA.Methods:A total of 12 RA patients and 11 age-matched healthy controls from University Hospital of Hubei University for Nationalities were recruited.Using lncRNA microarray technology to detect differently expressed lncRNAs in 3 cases of RA PBMCs and 3 cases of healthy controls.GO and Pathway analysis was performed.The coding-non-coding gene co-expression networks of lncRNA and mRNA was constructed based on the correlation analysis,and then searched lncRNA in pathogenesis of RA through the cis-analysis and trans-analysis.Results:A total of 1 615 deregulated lncRNAs and 878 deregulated mRNAs were detected in RA patients.GO analysis of different expressed mRNA may involve in metal ion binding,protein kinase binding,nucleotide binding,regulation of transcription,et al.Pathway analysis of different expressed mRNA may involve in TNF signaling pathway,B cell receptor signaling pathway,pancreatic cancer,system lupus erythematosus endometrial cancer,et al.REL,SMAD3 and ETS1 may play an important role in the pathogenesis and development of RA through cis-analysis and trans-analysis.As the NONHSAG027875,FR378506 and NONHSAT031501 also had the similar function,and they may be related to the pathogenesis and development of RA.Conclusion:Differentially expressed lncRNAs may exert a partial role in RA,and may provide potential targets for future treatment of RA.