1.Prediction,synthesis and characteristics of B-cell epitopes of CSFV envelope glycoprotein E2
Siguo LIU ; Gang MA ; Xinglong YU ; Maolin ZHANG ; Changchun TU ;
Chinese Journal of Immunology 2001;0(07):-
Objective:Study on characteristics of two synthesizd peptides based on CSFV E2 protein. Methods:B cell epitopes of CSFV E2 antigen were predicted using accessibility and flexibility schemes, associated with antigenicity , secondary structure and multiple sites prediction. Two antigen peptides (Pep1 and Pep2) have been designed and synthesized and their reactivety were detected with 8 McAbs and antiserum against mE2 protein, then the peptides were conjugated with BSA and immunized rabbits respectively. Results:Both Pep1 and Pep2 could react with antiserum and McAb A11, Pep2 could interact with McAbD5 and McAbD8. Only Pep1 BSA conjugate can stimulate high level and specific antibodies.Conclusion: The peptide1 has good antigenicity.
2.Effects of adenohypophysis function after treating with thyrotropin releasing hormone in patients with brain trauma
Bin LI ; Bao ZHAO ; Jingliang YE ; Feng YU ; Wenguo SHENG ; Guojun SU ; Qiang MA ; Xinglong XU
Chinese Journal of Biochemical Pharmaceutics 2014;(3):108-110
Objective To analyze the effects of adenohypophysis function after treating with thyrotropin releasing hormone and its clinical significance in patients with brain trauma. Methods There were 22 cases with traumatic brain injuries from July 2010 to September 2012 in Chinese people's Liberation Army nine eight hospital,after injuried within 4 to 12 hours,then 1 1 cases who were given thyrotropin-releasing hormone(TRH)were selected as experimental group,while 1 1 cases who were given the same amount of isotonic saline were selected as control group,then the score of GCS, ICS,RLS85 and the improvement of adenohypophysis function were observed. Results After treatment,the score of glasgow coma scale (GCS ), innsbruck coma scale (ICS),and the reaction level scale (RLS85)between two groups were significantly increased in three days compared with before treatment,and within three days after injury situation,the improvements of ICS and RLS85 in experimental group were better than control group(P<0.05 ). Compared with control group,the levels of each gland pituitary hormone in experimental group were significantly increased(P<0.05 ),and on the third day,the growth hormone (GH)was reduced significantly,finally 50%of that in control group. Conclusion Patients with brain injury treated with thyrotropin releasing hormone,has no significant adverse reactions,with the characteristics of safe and effective.
3.Rapid Detection of Porcine Circovirus 2 Based on Double Molecular Beacons
Chunyi TONG ; Bin JIANG ; Bin LIU ; Xinglong YU ; Wei WANG ; Wei LUO
Chinese Journal of Analytical Chemistry 2014;(8):1104-1109
A double-molecular beacons (DMB) based assay was developed for porcine circovirus 2(PCV2) detection. Two single-stranded DNA molecular beacons which could specifically hybridize with PCV2 genome DNA respectively in different sequence were designed according to the characteristics of the PCV2 genome sequences. The fluorescence signal was amplified 80 times by DMB, which was 2-4 times higher than that of single molecular beacon. Under the optimal conditions of 10 mmol/L MgCl2 , 20 mmol/L Tris-HCl (pH=8. 0), 40 ℃ and 30 min incubation time of DNA with DMB, the enlargement factor was increased linearly with DNA concentration over the range from 2 nmol/L to 200 nmol/L, with a detection limit of 1 nmol/L. The method was applied to detect PCV2 in genome of 18 swine fever samples and 8 PCV2 positive cases were found, which were confirmed by PCR method.
4.Effect of Mesalazine Hydroxyl-propyl-β-cyclodextrin Inclusion Compound Liquid Suppositories on Ulcera-tive Colitis in Rats
Shuhai YU ; Weizhong HAO ; Danggang YAO ; Peng LIU ; Lingli ZHANG ; Xinglong SONG
China Pharmacist 2016;19(10):1847-1849
Objective:To study the effect of mesalazine hydroxyl-propyl-β-cyclodextrin inclusion compound ( MSZ-HP-β-CD) liq-uid suppositories on ulcerative colitis ( UC) in rats. Methods:Dextran sulfate sodium was used to induce UC in rats. The marketed MSZ suppositories were applied as the positive control, and the changes in morphology and histopathology of the UC rats were observed after the treatment with MSZ-HP-β-CD liquid suppositories. Results:The results of morphology study showed that CMDI was decreased in the two MSZ preparation groups when compared with that in the model group (P<0. 01), and that in MSZ-HP-β-CD liquid supposi-tories group was the lowest (P<0. 05). The results of histopathology study showed that with the treatment of the marketed MSZ sup-positories and MSZ-HP-β-CD liquid suppositories, the ulcer and damage were improved, and the improvement in the MSZ-HP-β-CD liquid suppositories group was more significant. Conclusion: MSZ-HP-β-CD liquid suppositories exhibit better effect on UC in rats when compared with the marketed MSZ suppositories, which is worthy of further studies.
5.Effects of different doses of remifentanil on short-term learning and memory ability in the developing rats
Jian YANG ; Zilong YU ; Hong GAO ; Xiaohua ZOU ; Wei OU ; Xinglong XIONG
The Journal of Clinical Anesthesiology 2016;32(9):910-913
Objective To investigate the effects of different doses of remifentanil on learning and memory ability,the expression of hippocampal tissue phosphorylation of cAMP response element binding protein (p-CREB)in developing rats.Methods A total of 72 Sprague-Dawley rats (1 9-23 g) were randomly divided into 4 groups (n =18 each):Group C:normal saline control group;R1,R2, R3 group received continuous intraperitoneal remifentanil 1,5,10 μg·kg-1 ·min-1 for 2 hours re-spectively.Both total volume of remifentanil and saline were 2 ml.The SpO 2 and pulse rates were mo-nitored during the experiment.Step-down test was used to evaluate the learning and memory ability, while Western blot analysis was performed to measure the expression of hippocampal p-CREB protein in 4 h,24 h,1 week when the rats were awake.Results Compared with group C,group R1 and R2, pulse rates of group R3 decreased significantly (P <0.05 ),but the changes of SpO 2 in each group were not statistically significant.At 4 h point:compared with group C and group R1,the error times in step-down test were increased in both group R2 and R3,the latencies were shortened (P <0.05);Compared with group R2,the error times were increased in group R3,latency was shortened (P <0.05).At 24 h point,compared with group C and group R1,the error times were increased in group R2,R3,latencies were shortened (P < 0.05 );Compared with group R2,the error times were in-creased in group R3,latency was shortened (P <0.05 ).The error times and latency of each group had not statistical significance in one week.At 4 h point,the expression of p-CREB protein in hippo-campus of group R3 downregulated compared with group C and group R1,R2,respectively (P <0.05).At 24 h point,the expression of p-CREB protein in hippocampus of group R2,R3 decreased compared with group C and group R1 respectively(P <0.05);The expression of p-CREB protein in each group had no statistical significance in one week.Conclusion 5-10 μg · kg-1 · min-1 dose of remifentanil can result in a decline of learning and memory ability in the developing rats in short-term,and the mechanism may relate to the inhibition of p-CREB protein expression in hippocampus.
6.Craniocerebral injury promotes sciatic nerve regeneration
Xinze HE ; Wei WANG ; Jianjun MA ; Tiemin HU ; Changyu YU ; Yunfeng GAO ; Xinglong CHENG ; Pei WANG
Chinese Journal of Tissue Engineering Research 2016;20(27):4061-4067
BACKGROUND:Studies have shown that craniocerebral injury can promote the repair of sciatic nerve injury in rats, but its precise mechanism remains unclear.
OBJECTIVE:To further explore the action mechanism of craniocerebral injury on the repair of sciatic nerve injury using morphology and histology.
METHODS:Sixty specific-pathogen-free healthy male Sprague-Dawley rats were randomly divided into two groups. Rats with craniocerebral injury and sciatic nerve injury were considered as the experimental group. Rats with simple sciatic nerve injury were considered as the control group. Classical Feeney method was used in models of craniocerebral injury and SunderlandV sciatic nerve injury. At 8 and 12 weeks after modeling, sciatic nerve index was detected. Masson staining and NF200 immunofluorescence staining were used to observethe nerve regeneration atthe anstomotic site. Transmission electron microscope was used to observe the number of regenerative axons.
RESULTS AND CONCLUSION:At 8 and 12 weeks after modeling, compared with the control group, gait and sciatic nerve index recovered better in the experimental group. In the experimental group, Masson staining showed fewer nerve membrane colagen fibers, and the axon arranged neatly.NF200 immunohistochemistry showed that in the experimental group, the density of regenerated nerves was high, and nerveswere regularly distributed. Transmission electron microscopy showed that in the experimental group, regenerative axons were regularly arranged, colagen scar was less, and myelin layer arranged regularly. Results suggested that the craniocerebral injury in rats may promote the repair of peripheral nerve injury by reducing scar colagen in nerve endings.
7.Peripheral nerve repair:theory and technology application
Xinze HE ; Wei WANG ; Tiemin HU ; Jianjun MA ; Changyu YU ; Yunfeng GAO ; Xinglong CHENG ; Pei WANG
Chinese Journal of Tissue Engineering Research 2016;20(7):1044-1050
BACKGROUND:Recovery of motor and sensory function from peripheral nerve injury is relatively slow and incomplete. It is a difficult problem for orthopedic surgeons that mainly leads to the decline in the quality of life in patients.
OBJECTIVE: To conclude the methods and corresponding outcomes in peripheral nerve regeneration by analyzing the new treatment means for peripheral nerve injury.
METHODS:PubMed, Wanfang, CNKI databases were retrieved for relevant articles using key words of “nerve injury, regeneration”, and then retrieval data were sorted and analyzed.
RESULTS AND CONCLUSION:In recent years, in-depth studies on peripheral nerve repair have been made in the folowing aspects: surgical mode, drug, cytokine, gene transfer and biomaterials as wel as traditional Chinese medicine. If the detect size is four times longer than the diameter of nerves, the nerve regeneration chamber can achieve good outcomes. The methods of restoring nerve continuity folowing nerve injury are developed from surgical anastomosis to photochemohistological method, thermal laser welding, plastic repair and other emerging technologies. Studies have found that plasminogen activator, nerve growth factor, neurotrophic factor, recombinant erythropoietin, human tissue kalikrein, B vitamins and their derivatives, herbal preparations, immunosuppressive agents al can promote nerve regeneration.
8.Analysis and comparison strategy of mixed DNA profile without known provider
Yu WANG ; Kunyun MAO ; Jiajia CHEN ; Xinglong HAO ; Run JIA
Chinese Journal of Forensic Medicine 2017;32(6):645-648
Objective From the perspective of making full use of database comparison function, giving certain guidelines to analyze mixed DNA profile,compare database,screen comparison results. Methods Using CPI to describe the identification of mixed DNA profile.Using CPBI to estimate reliability of individual samples being included. Results When CPI is less than 10-7, mixed DNA Profile is worth to be compared in database.When the number of alleles at one locus is more than 2, retain an additional allele will not reduce identification too much. According to the CPBI of the included samples,we can find the most reliable sample.
9.Fusion expression of Escherichia coli heat-labile enterotoxin B subunit gene and foot-and-mouth disease virus type O VP1 gene and immunogenicity analysis.
Runcheng LI ; Xinglong YU ; Xia BAI ; Weijun XIANG ; Meng GE ; Manxiang LI
Chinese Journal of Biotechnology 2009;25(4):560-565
LTB gene fragment was amplified by PCR from plasmid pMDTLT, and a recombinant plasmid pETLTBVP1 was constructed by inserting LTB gene fragment into VP1 gene expression plasmid pETVP1 constructed previously. The recombinant plasmids were transformed into E. coli BL21(DE3) and induced to express by IPTG. The recombinant protein existed in the inclusion body and its molecular weight was about 39 kD proved by SDS-PAGE analysis. Western blotting showed that the fusion protein could be reacted with both anti-FMDV and anti-cholera toxin serum demonstrating the immunoactivity of the fusion protein. Strong immune responses can be induced in mice inoculated with the fusion protein intraperitoneally, and the serum antibody level is higher than that of commercial foot-and-mouth disease vaccines.
Animals
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Antibodies, Viral
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blood
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Bacterial Toxins
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genetics
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immunology
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metabolism
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Capsid Proteins
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genetics
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immunology
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metabolism
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Enterotoxins
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genetics
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immunology
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metabolism
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Escherichia coli
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genetics
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metabolism
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Escherichia coli Proteins
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Female
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Gene Fusion
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genetics
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Mice
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Plasmids
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genetics
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Recombinant Fusion Proteins
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genetics
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immunology
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metabolism
10.Rapid detection of Pseudomonas aernginosa by the fluorescence quantitative TaqMan PCR assay targetting ETA gene.
Xinglong XIAO ; Jingwei ZHANG ; Jun GONG ; Yanping PAN ; Yigang YU ; Xiaoquan YANG ; Hui WU
Chinese Journal of Biotechnology 2008;24(4):581-585
Pseudomonas aernginosa (PA) is one of the most universal pathogens in clinical diagnosis, and conventional detection assay has many disadvantages. In this research, a pair of specific primers and a TaqMan fluorescent probe were designed in the conservative region of ETA gene by the method of bioinformatics analysis, the detection method for PA was successfully developed. Different gradient concentrations of PA DNA and various pathogen DNA were amplified by fluorescence quantitative PCR (FQ-PCR) to confirm the specificity and sensitivity of the developed method. Results showed that the developed detection assay is more sensible and specific by comparison to the conventional FQ-PCR method, and it is valuable for research and application prospects.
ADP Ribose Transferases
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genetics
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Bacterial Toxins
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genetics
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DNA, Bacterial
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analysis
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Exotoxins
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genetics
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Fluorescent Dyes
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Fluorometry
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methods
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Polymerase Chain Reaction
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methods
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Pseudomonas aeruginosa
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genetics
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isolation & purification
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Sensitivity and Specificity
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Taq Polymerase
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Virulence Factors
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genetics