2.Screening and characterization of aptamers of Cepsilon3-Cepsilon4 protein.
Zhongcheng LIU ; Lijun ZHAO ; Yanfen ZHANG ; Hailang SHI ; Yao XIE
Acta Pharmaceutica Sinica 2012;47(12):1605-11
In order to obtain nucleotides aptamers bind to IgE, 80 bp nucleotides single-stranded DNA library containing 40 random nucleotides was designed and synthesized. Oligonucleotides that bind to human Cepsilon3-Cepsilon4 protein were isolated from ssDNA pools by the systematic evolution of ligands by exponential enrichment (SELEX) method using nitrocellulose filters as screening medium. Through the optimization of critical PCR and asymmetric PCR parameters including annealing temperature, cycles, and molar ratios of target protein and ssDNA etc, a suitable screening system was established. The aptamers of Cepsilon3-Cepsilon4 protein with high affinity and high specificity were identified by ELISA with biotin-streptavidin-horseradish peroxidase system, and its primary sequence and second structure were analyzed by DNAMAN package and DNA folding sever after being cloned and sequenced. Moreover, target protein was bound to one aptamer and another aptamer modified with biotion together forming a sandwich-like complex, which was captured in microwell to detect IgE concentration using the optimal combination in the sandwich method named enzyme-linked aptamers sorption assay (ELASA). The method could be used for the quantitative detection of human IgE, and whose sensitivity reached to 120 ng x mL(-1).
3.Expression and signiifcance of PLTP and MIF in mice with bronchopulmonary dysplasia
Guifang LI ; Baohai SHI ; Guo YAO ; Junli LIU ; Chunhua QI
Journal of Clinical Pediatrics 2013;(9):876-879
Objective To study the expression and signiifcance of phospholipid transfer protein (PLTP) and macrophage migration inhibitory factor (MIF) in mice with bronchopulmonary dysplasia (BPD). Methods Ninety-six 4-day-old mice were randomly divided into oxygen group and air group. Mice in oxygen group were exposed to a FiO2 of 65%, and mice in air group were exposed to air. On day 7, 14, 21 and 28, blood and lung tissue samples from 12 randomly selected mice in each group were obtained. The serum levels of MIF and PLTP were measured by ELISA assay. The morphological changes of lung tissue were ob-served with HE staining. Results The mice in oxygen group showed thickened lung parenchyma and obvious pulmonary ifbrosis. The radioactive alveolar count was signiifcantly lower in oxygen group than that in air group (P<0.01). PLTP level in air group was increased gradually from day 7 to day 21, and began to decrease on day 28. PLTP level in oxygen group was increased from day 7 to day 14, and decreased on day 21 and day 28. MIF level in air group did not change during the experiment. MIF level in oxygen group was signiifcantly increased from day 7 to day 21, and began to decrease on day 28. Conclusions MIF and PLTP may be good biomarkers for the diagnosis of BPD.
5.Role of adipokines in the occurrence and regulation of autoimmune diseases
Yang LIU ; Rui YANG ; Hongbin YUAN ; Jiangang SHI ; Dingkang YAO
Chinese Journal of Tissue Engineering Research 2017;21(16):2612-2618
BACKGROUND: Adipokines are secreted by adipose tissue, including a series of cytokines and chemokines. Additionally, adipokines have been found to play certain roles in pro- and anti-inflammatory regulation. OBJECTIVE: To analyze the regulatory effect of various adipokines in the inflammatory process of autoimmune response and immune diseases, thus providing the strategy for controlling and treating autoimmune diseases and searching effective indexes for assessing disease development, incidence of complications and prognosis. METHODS: CNKI and PubMed databases were retrieved for the articles addressing the role of adipokines in autoimmune diseases published from 1986 to 2016 using the keywords of adipokine, autoimmune in English and Chinese, respectively. RESULTS AND CONCLUSION: Adipose tissue plays a critical role in metabolism regulation, and its effect on the immune system has emerged through a series of studies concerning adipokines regulating immune response. Adipokines exert various functions, and regulate metabolism and inflammation. In the development and occurrence of autoimmune diseases, some factors for immune responses remain unclear, and in-depth studies on adipokines will be helpful for lifting the veil of autoimmune diseases.
6.Investigation of Human Papillomavirus-16 Infections and Its Mixed Infection in Yunnan Region
Zheng LI ; Si CHENG ; Lei SHI ; Yufeng YAO ; Feng LIU
Journal of Kunming Medical University 2014;(1):92-94
Objective To investigate the distribution of Human Papillomavirus 16 (HPV16) infection and its mixed infection with other HPV subtypes in the Yunnan region. Methods 16 166 cases of women were tested using flow fluorescence Luminex technology. Results (1) HPV16 infection rate and mixed infection rate was 2.2%and 28.0%, respectively; (2) The most common type of HPV16 mixed infection was HPV52, followed by HPV33. The two kinds of mixed infection accounted for 39.8% of the total infection rate; (3) There was a significant difference between each age group of HPV16 mixed infection (Chi=26.39, <0.01) . Conclusion The HPV16 infection was mainly HPV infection in Yunnan region. HPV16 mixed infection merged mainly with HPV52 and HPV33. HPV16 mixed infection was associated with age.
7.Disinfection of Dental Hand Piece Contaminated with HBV in Practice
Guangliang NIU ; Dong ZENG ; Gang LIU ; Lei SHI ; Yao XIE
Chinese Journal of Nosocomiology 2006;0(06):-
OBJECTIVE To investigate the disinfection efficacy of dental high-speed hand-pieces contaminated by HBV with the method of pressure steam sterilizer.METHODS Taking 50 oral clinical patients randomly,and three group-samples which included saliva,hand-pieces contaminated by clinical operation and disinfected by pressure steam sterilizing were collected,then the samples were detected HBsAg and HBV DNA,respectively.RESULTS There were 95% of the saliva samples being HBsAg positive,the positive rate of the high-speed hand-pieces contaminated by clinical operation was lower than the saliva samples,and the positive rate of the hand-piece disinfected by pressure steam sterilizing was the lowest.HBV DNA was undetectable in the sample before or after disinfected hand-pieces used in patients′ saliva which HBsAg s/co value higher than 5.0.CONCLUSIONS Pressure steam sterilizing is effective to reduce the contaminated HBV on hand-pieces,but the biology test should be taken to demonstrate whether the complete sterilizing is achieved.
8.A comparative study on vascular blockade during hemihepatectomy for primary hepatic carcinoma
Zheng SHI ; Mingzhi YANG ; Jingfeng LIU ; Jieyin GONG ; Zhixiong YAO
Chinese Journal of General Surgery 2000;0(11):-
Objective To compare among three different methods in performing hepatic vascular blockade during hemihepatectomy for primary hepatic carcinoma ( PHC). Methods Between 2000 and 2005, 83 PHC patients underwent the Pringle's maneuver (Group A) , 67 cases in combination Pringle's maneuver and inferior vena cava (IVC) clamping (Group B) , and 32 cases received anatomical blood flow blockade for the half liver for hemihepatectomy ( Group C). Results Operation time in group C was longer than that in group A and B(t =3. 27、2. 74,all P
9.Application of Intra-Operative Cholangiography
Xing-yao, SHI ; Chao-hui, LIU ; Lin-xing, LU
Journal of Shanghai Jiaotong University(Medical Science) 2001;21(2):182-183
Objective To evaluate the application of intra-operative cholangiography in cholecystectomy. Methods A total of 442 patients with biliary calculi underwent cholecystotomy were reviewed. Results Of the 442 cases, 3 were false positive and 1 was false negative radiologically. The accuracy rate was 99.1%. Conclusion The results suggested that the application of intra-operative cholangiography could decrease the incidence of residual calculi and avoid unnessary exploration and trauma of the common bile duct.
10.Mechanisms and Effect of Ginkgolide B on the Differentiation of Neuron Stem Cells
Yonghong WANG ; Xue LUO ; Yongjiang SHI ; Hongliang LIU ; Zhongxiang YAO
Chinese Journal of Rehabilitation Theory and Practice 2007;13(8):701-703
Objective To observe the effect of Ginkgolide B of various consistency on the differentiation of neuron stem cells (NSCs).MethodsNSCs were cultured in differentiation medium containing Ginkgolide B of various consistency for 3 and 7 days, the neurites length and cell body area were measured by inverted phase-contrast micrograph, then neurofilament-200 (NF-200), glial fibrillary acidic protein (GFAP), adenomatus polyposis coli (CC-1) expression were detected and counted by fluorescence microscope. The suppressor of cytokine signaling-2 (SOCS2), inhibitor of DNA binding-2 (Id2) were alsoimmunostained. The percentage of positive cells were counted respectively.ResultsThe neurites length and cell body area in Ginkgolide B groups were obviously larger than that in the control group. The percentage of NF, GFAP positive cells in Ginkgolide B groups increased with dosage increasing of Ginkgolide B. Compared with the normal control group, the percentage of SOCS2 positive cells increased significantly ( P<0.01) and the percentage of Id2 positive cells decreased significantly ( P<0.01) in Ginkgolide B groups.ConclusionGinkgolide B can promote NSCs to differentiate into neuron and astrocyte, the percentage of astrocyte is increased with a dosage-dependent relationship with Ginkgolide B.