3.Analysis of Leptospira interrogans ompA gene and immunological identification of its recombinant expression product
Wei DING ; Haiyan DONG ; Feng XUE ; Jie YAN ; Yongliang LOU
Chinese Journal of Microbiology and Immunology 2009;29(4):370-374-
Objective To investigate the distribution of ompA gnne in 15 Chinese reference standard strains belonging to 15 serogroups of Leptospira interrogate, and to express recombinant OmpA ( rOmpA ) and to identify immunogenicity and immunoprotection of rOmpA. Methods Genomic DNAs from different leptospiral strains were extracted by phenol-chloroform method. Entire ompA gene fragments from the strains were amplified by PCR and then sequenced after T-A cloning. A prokaryotic expression system of ompA gene from L. interrogans strain 56601 was constructed, and the expression and yield of rOmpA were determined by SOS-PAGE plus Bio-Rad Agarose Image Analyser. Rabbits were immunized with rOmpA for obtaining antiserum, and immunodiffusion test was used to measure the antiserum's titer. Western blot assay was performed to determine the immunoreaetivity of rOmpA with the antiserum against rOmpA and antiserum against whole cell of L. interrogans strain 56601, while mi-croscopic agglutination test (MAT) was applied to detect the cross agglutination to the 15 L. interrogans strains. A leptospire adhering cell model and a leptospire infecting guinea pigs model were used to determine the adhesion-bloc-king effect of rOmpA antiserum and immunoprotection of rOmpA. Results All the 15 L. interrogans strains, but not L. biflexa strain Patoe Ⅰ , had sequence conserved ompA genes. The yield of rOmpA was approximate 20% of the total bacterial proteins, rOmpA could induce rabbits to produce antibody and immunodiffusion titer of the anti-serum was 1:4. Both antisera against rOmpA and against whole cell of L. interrogans strain 56601 were able to pro-duce positive Western blot signs to rOmpA, and the former offered 1 : 20-1 : 320 MAT titers to the 15 L. interrogans strains. 1: 10-1:160 dilutions of rOmpA antiserum could efficiently block L. interrogans strain 56601 adhering to J774A. 1 cells, and 100 μg and 200 μg rOmpA displayed 50.0% and 75.0% immunoprotective rates in the infee-ted guinea pigs. Conclusion ompA gene only exists in genomes of different pathogenic L. interrogans serogroups. rOmpA has relatively stronger antigenicity, cross immunoreactivity and certain immunoprotection, implying that this recombinant protein may be used as a candidate antigen for developing universal genetic engineering vaccine of L. interrogans.
4.Changes of Cytokines in Newborn Meconinum Aspiration Syndrome
ning, DING ; hong-bing, WANG ; xue-ping, DONG
Journal of Applied Clinical Pediatrics 2006;0(22):-
Objective To investigate role of interleukin-1?(IL-1?),interleukin-8(IL-8),tumor necrosis factor-?((TNF-?)) and insulinlike growth factor-1(IGF-1) in meconium aspiration syndrome(MAS) in newborns.Methods The concentrations of(IL-1?),IL-8,TNF-? and IGF-1 in peripheral serum were detected in 32 newborns with MAS and 15 normal newborns by radio-immunity at the first,the third and seventh day after being born.Correlations between IGF-1 and changes of IL-1?,IL-8,TNF-? was analyzed.Results Compared with control group,the concentration of IL-1?,IL-8 and TNF-? increased significantly in newborns with MAS in three days after being born(all P0.05).On the other hand, the concentration of IGF-1 in newborns with MAS was obviously lower than that of normal newborns one week after birth.The concentration of IGF-1 showed negative correlations with that of IL-1?,IL-8 and TNF-?(r=-0.67,-0.61,-0.73 all P
5.Status quo and influencing factors of hyperlipidemia management in patients with contracted family doctor service
Beibei SONG ; Ai FENG ; Yuming DONG ; Jing DING ; Xue CHEN
Chinese Journal of General Practitioners 2021;20(3):321-326
Objective:To evaluate the status quo and influencing factors of hyperlipidemia management in patients with contracted family doctor service in the community.Method:The baseline data and blood lipid testing results of 752 hyperlipidemia patients (334 males and 418 females) with contracted family doctor service in Yuetan Community Health Service Center from November?2019 to May 2020 were collected. The hyperlipidemic patients were managed by family doctors based on atherosclerotic cardiovascular diseases(ASCVD) riks assessment. The ASCVD risk levels and low-density lipoprotein cholesterol (LDL-C) compliance rate of patients with different general data were compared, and the influencing factors of LDL-C control failure were analyzed by logistic regression.Results:The ASCVD risk assessment showed that among 752 patients there were 172 cases of low risk(22.87%), 167 cases of moderate risk(22.21%),352 cases of high risk(46.81%) and 61 cases of extremely high risk(8.11%). A significant difference was detected in sex,rate of smoking,incidence of overweight or obesity among patients with different ASCVD risk levels ( P<0.05).The overall control rate of LDL-C was 48.8% (367/752), that for low, moderate, high and extremely high risk patients were 83.73% (144/172), 53.89% (90/167), 34.38% (121/352) and 19.67%(12/61), respectively. A significant difference was detected in sex(female: 52.87%, 221/418),age(aged over 80: 58.82%, 110/187), rate of smoking (non-smoking:52.40%, 327/624) and medication compliance (good compliance:52.87%,221/418) between LDL-C control and uncontrol groups (χ2=6.323,11.816,19.022,25.274; P<0.05). Multiple logistic regression analysis revealed that male gender ( OR=1.800,95% CI:1.325-2.419), smoking ( OR=2.630,95% CI:1.726-4.007) and poor medication compliance ( OR= 2.179, 95% CI: 1.581-3.003) were independent risk factors for uncontrolled LDL-C levels. Conclusion:Patients with hyperlipidemia have a relatively high risk of cardiovascular diseases, and their blood lipids are not well controlled. The management of blood lipid should be enhanced in patients with chronic diseases, particularly for male patients with smoking and poor medication compliance.
6.The application of septum cartilage in correcting of the unilateral cleft lip nasal deformity.
Lipeng DONG ; Chunling XUE ; Fang DONG ; Yuhu DING ; Rongqiang LI
Chinese Journal of Plastic Surgery 2002;18(1):15-17
OBJECTIVETo explore an ideal method for comprehensive correcting of the unilateral cleft lip nasal defomity.
METHODSThe operation includes replacement of the displaced tissue, excision of the deviated septum, and correction of the deviated septum and the deformity of the lower nose. The excised nasal septum was grafted on the alar base, the nasal columella and the nasal tip.
RESULTS32 patients were treated with this method from 1994 to 1999. Postoperative fellow-up for 1-3 years demonstrated satisfactory results.
CONCLUSIONThrough excision and grafting of the nasal septum, the deviated septum and the deformity of the lower nose are effectively corrected. Complete undermining and replacement of the displaced structure are the basis of correcting the cleft lip nasal deformity.
Adolescent ; Adult ; Cartilage ; transplantation ; Cleft Lip ; surgery ; Female ; Humans ; Male ; Nasal Septum ; transplantation ; Nose ; abnormalities ; surgery
7.LC-MS/MS assay of methylphenidate: stability and pharmacokinetics in human.
Xue-Mei LUO ; Li DING ; Xin GU ; Li-Yuan JIANG ; Xin DONG
Acta Pharmaceutica Sinica 2014;49(1):83-88
The study aims to solve the instability problem of methylphenidate (MPH) in plasma, and establish a LC-MS/MS method for simultaneous determining of MPH in human plasma. The stabilities of MPH in different media were studied, and the degradation characteristics of MPH in these media were also investigated by HPLC and LC-MS/MS. To a 200 microL aliquot of freshly collected plasma sample, 10 microL 2% formic acid was added immediately to prevent the hydrolysis of MPH in human plasma samples. Chromatographic separation was performed on a Sapphire C18 column using the mobile phase of methanol - 5 mmol.L-1 ammonium acetate buffer solution containing 0.1% formic acid (46 : 54). MPH was quantified by tandem mass spectrometry operating in positive electrospray ionization mode with multiple reaction monitoring. The detection used the transitions of protonated molecules at m/z 234.2-->84.1 for MPH and m/z 260.3-->183.1 for propranolol (IS), separately. The intra- and inter-assay precisions were all below 5.0%. The accuracies were all in standard ranges. The linear calibration curve was obtained in the concentration range of 0.035-40 ng.mL-1. The methods fulfilled the demand. The method was used to determine the concentration of MPH in human plasma after a single dose of 36 mg MPH tablet to 6 healthy Chinese volunteers. The method is suitable for the precisely determination of MPH and for pharmacokinetic study of MPH in human plasma.
Adult
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Central Nervous System Stimulants
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blood
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pharmacokinetics
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Chromatography, High Pressure Liquid
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Chromatography, Liquid
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Drug Stability
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Humans
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Male
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Methylphenidate
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blood
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pharmacokinetics
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Spectrometry, Mass, Electrospray Ionization
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Tandem Mass Spectrometry
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Young Adult
9.Changes of S-100? and Glial Fibrillary Acidic Protein in Serum of Children with Epilepsy
li, GAO ; ying-xue, DING ; yan-ping, LIU ; xuan, ZHANG ; juan, LIU ; dong-ju, MA
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To explore the change of S-100? and glial fibrillary acidic protein(GFAP)in serum after seizure and medication in children with epilepsy.Methods Serum protein level of S-100? and GFAP were determined by double antibody sandwish enzyme-linked immunosorbent assay(ELISA)in 41 cases with epilepsy and 30 healthy children.The specimen of venous blood were taken by 24 hours after seizure,4 weeks,12 weeks after medicine and their supernate preserved at-80 ℃ after centrifugat.Results Twenty-four hours after seizure,protein level of S-100?,GFAP in serum was significantly higher than that of control group(Pa0.05).Four weeks after medication,protein level of S-100?,GFAP in serum of epileptic group decreased,but still higher than that in control group,and the difference was significant(P
10.Strategies of endodontic infection control.
Ding-ming HUANG ; Xue-dong ZHOU
West China Journal of Stomatology 2011;29(3):225-228
Microbe infection is not only the key pathogenic factor of primary endodontic infectious diseases, but also the arch-crimina of endodontic treatment fail. Therefore, the endodontic treatment success is based on the debridement of infectious root canal, elimination of bacteria, and the prevention of endodontic reinfection. It is beset with difficulties to control the endodontic infection in clinic because of the bacterial variety, anatomic complexity of root canal, and limitation of the root canal therapeutic methods. In order to get the ideal prognosis of the root canal treatment, in the procedure of the therapy, it should be the tight isolation of tooth, reasonable choose of the root canal preparation technique, rational use of root canal preparation instrument, effective irrigation of root canal, proper root canal dressing, and the 3-dimensional obturation of root canal.
Humans
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Infection Control
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Root Canal Irrigants
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Root Canal Preparation
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Root Canal Therapy