1.Advances in the molecular genetics of the ankylosing spondylitis
International Journal of Pediatrics 2012;(6):588-590
Ankylosing spondylitis (AS) is a chronic rheumatic disease involving the spine and sacral hip.AS is the prototype disease of seronegative arthritis of the spine.Besides the violations of the spine,AS can also lead to violations of the peripheral joints,even the eyes,intestine,aortic root,and other tissues and organs.The pathogenesis of AS has remained unclear.AS,a genetic disease,is influenced by the genetic and environmental factors.Studies have suggested that HLA-B27 is the most critical genes to AS,and forming three kinds of hypotheses associated with HLA-B27.This paper reviews the molecular genetics and pathogenesis of AS.
2.Study on genetic epidemic of ankylosing spondylitis
Junjie LI ; Jiafen JI ; Changyun LIU
International Journal of Pediatrics 2013;40(4):413-415
Objective To investigate the epidemiological situation and the mode of inheritance of ankylosing spondylitis (AS).Methods The AS prevalence investigation was carried out from three different areas in Shandong Province to investigate the incidence of the 80 patients' family members of sick group and calculate the heritability of AS.Results After the investigation and calculation we found the incidence of primary relatives was 7.23%,the heritability was 85.94%,the incidence of secondary relatives was 0.94% and the heritability was 57.4%,the incidence of thirdly relatives was 0.39%,the heritability is 36.2%,the average heritability among primary,secondary and thirdly was 68.47% ± 1.64%.Separation ratio was 0.088 9.Conclusion AS is a polygenic disease,its average heritability was 68.47% ± 1.64% and separation ratio is 0.088 9.By this research,the incidence risk of AS patients' pedigree offspring can be assessed,then strengthen protective factors to reduce the incidence of the disease to provide the basis for eugenics.
3.Associations of -2548A/G polymorphism in leptin gene and Gln223Arg polymorphism in leptin receptor gene with asthma and metabolic syndrome
Yunxiao LI ; Xia JI ; Junjie GAO
Chinese Journal of Immunology 2014;(11):1467-1471,1476
Objective:To investigate the relationship between polymorphism of leptin gene -2548A/G and leptin receptor gene ( LEPR) Gln223Arg and asthma and metabolic syndrome.Methods: 82 asthma-combined metabolic syndrome patients,114 asthma patients,100 metabolic syndrome patients and 96 normal controls were conducted.The polymerase chain reaction and restriction fragment length polymorphism ( PCR-RFLP) analysis were performed to investigate the polymorphism of leptin gene -2548A/G and LEPR gene Gln223Arg site.In addition, asthma was graded into mild AS and mod-severe AS according to lung function.Then the associations between the polymorphism of leptin gene -2548A/G and LEPR gene Gln223Arg and different grades of asthma were performed.Results:①The biochemical indicators were different compared between each group.②The genotype and allele frequencies in leptin gene polymorphism -2548A/G were significantly difference between metabolic syndrome patients(P=0.047 and 0.046), asthma-combined metabolic syndrome patients( P=0.038 and 0.044) ,mod-severe asthma patients( P=0.019 and 0.028) and control group.③There was a significant difference of genotype and allele frequencies in LEPR gene Gln223Arg between metabolic syndrome patients and controls(P=0.037 and 0.023);between metabolic syndrome patients and asthma patients(P=0.000 and 0.000).There was a significant difference of allele frequencies in LEPR gene Gln223Arg between asthma-combined metabolic syndrome patients and asthma(P=0.032) .Conclusion: Polymorphisms of the leptin gene -2548A/G site may be associated with metabolic syndrome and mod-severe asthma.Polymorphisms of the LEPR gene Gln223Arg site may be only associated with metabolic syndrome.The two genes would be the candidate genes in early prevention and control.
4.HUMAN TUMOR VACCINE GENERATED BY FUSION OF HEPATOCELLULAR CARCINOMA CELLS WITH ACTIVATED B CELLS AND ITS FUSION RATE DETECTION
Congxin ZHANG ; Junjie JI ; Weizh QIAN
Medical Journal of Chinese People's Liberation Army 2001;0(09):-
Fresh hepatocarcinoma tissue and spleen samples were taken from patients during surgery. B cells from spleen were purified and activated. The hepatocarcinoma vaccine was made by cell fusion between hepatic tumor cells and activated B cells. PEG was used as the fusion agent. The fusion cells were cultured and deactivated. MHC Ⅱ and B7 molecules on activated B cells were determined by flow cytometry. Fusion rate and recovery rate of cells after refrigeration were determined respectively at the same time. The results showed that MHC Ⅱ and B7 molecules on the activated B cells were enhanced comparing with B cell. The fusion rates of three cases were 66 84%, 74 43%, and 76 55%, respectively. The recovery rates of cells were 95% and 97% after DMSO and glycerol refrigeration, respectively. The results suggest that the hepatocarcinoma vaccine owns high fusion rate and recovery rate of cells after refrigeration. It's easy to make and store. So the hepatocarcinoma vaccine is suitable for clinical use.
5.Recombinant human brain myelin basic protein and its antibody preparation
Ji LIU ; Ruohan WANG ; Yu LIU ; Junjie CHEN
Chinese Journal of Tissue Engineering Research 2006;10(21):170-171
BACKGROUND: Central nervous system (CNS) myelin is made up of 70% lipids and 30% proteins with human brain myelin basic protein (hMBP) constituting 1/3 of the proteins. MBP is a family of proteinswith four isoforms only in human brain. OBJECTIVE: To investigate the expression of MBP and its immunological function. DESIGN: Single sample study. SETTING: Department of Biochemistry and Molecule Biology, Huaxi College of Priclinical Medicine and Forensic Medicine, Sichuan University. MATERIALS: This experiment was conducted at Department of Bio chemistry and Molecule Biology, Huaxi College of Priclinical Medicine and Forensic Medicine, Sichuan University from August 2003 to March 2004. Relative molecular weight was 215000 hMBP cDNA clone pGEMP,prokaryotic expression recombinant vector pGEX-5T, T4 DNA Ligase,calf intestine alkaline phosphates, X-gal, IPTG, 123 Ladder (BRL), nitrocellu lose ,4-chloro-1-Naphthol;MBP and Anti- MBP ELISA kit. INTERVENTIONS: ①hBMP gene cDNA clone segment was digested with EcoR1 and BamH1; ②Construction and transformation of the recombinant expression vector p5TMP; ③ The growth and induced expression of the recombinant expression vector transformed bacteria; ④ Preparation of the antibody of recombinant hMBP. MAIN OUTCOME MEASURES: ① Detection and identification of the expressed protein product; ② Detection and identification of hMBP anti body. RESULTS: ① Screening and identification of recombinant MBP: The 4 999 bp pGEX-5T DNA and 557 bp MBP inserted fragment were obtained, indicating that the white colonies contained recombinant expression plasmid of exogenous DNA; ② A dense band disappeared from the control plasmid while a new special polypeptide band with apparent molecular weight 42 000 was detected in recombinant cell lysate; ③After 5 subcutaneous injections, antibody was obtained at 1:16 titer. The specificity of the antibody to MBP was confirmed.CONCLUSION: Compared with the original expression vector, the new constructed expression vector containing 215 000 MBP exons Ⅰ-Ⅶ coding sequence was not only without the coding area defects of 34 bp in 5' sequence but also expressed more highly in prokaryote obviously. In addition, the recombinant MBP antibody prepared successfully.
6.Analysis of the results of bilingual teaching with seven-year medical students
Fang WANG ; Lin ZHANG ; Yanjin MOU ; Junjie TAO ; Xian JI
Chinese Journal of Medical Education Research 2011;10(2):183-185
By analyzing data from a questionnaire survey on bilingual teaching, we evalueted the results of bilingual teaching for seven-year medical students and the main problems of bilingual teaching and solutions accordingly. An uneven English level on the part of teachers and students and imperfect materials etc. affect the overall results of bilingual teaching. Therefore persistent efforts need to be made in enhancing the teachers' English level, improving teaching methods and compiling proper textbooks so as to genuinely improve the bilingual teaching program.
7.Evaluation of penicillin expandase mutants and complex substrate inhibition characteristics at high concentrations of penicillin G.
Linjun WU ; Keqiang FAN ; Junjie JI ; Keqian YANG
Chinese Journal of Biotechnology 2015;31(12):1690-1699
Penicillin expandase, also known as deacetoxycephalosporin C synthase (DAOCS), is an essential enzyme involved in cephalosporin C biosynthesis. To evaluate the catalytic behaviors of penicillin expandase under high penicillin G concentration and to identify mutants suitable for industrial applications, the specific activities of wild-type DAOCS and several mutants with increased activities toward penicillin G were determined by HPLC under high penicillin G concentrations. Their specific activity profiles were compared with theoretical predictions by different catalytic dynamics models. We evaluated the specific activities of wild-type DAOCS and previous reported high-activity mutants H4, H5, H6 and H7 at concentrations ranging from 5.6 to 500 mmol/L penicillin G. The specific activities of wild-type DAOCS and mutant H4 increased as penicillin G concentration increased, but decreased when concentrations of substrate go above 200 mmol/L. Other mutants H5, H6 and H7 showed more complex behaviors under high concentration of penicillin G. Among all tested enzymes, mutant H6 showed the highest activity when concentration of penicillin G is above 100 mmol/L. Our results revealed that the substrate inhibition to wild-type DAOCS' by penicillin G is noncompetitive. Other DAOCS mutants showed more complex trends in their specific activities at high concentration of penicillin G (>100 mmol/L), indicating more complex substrate inhibition mechanism might exist. The substrate inhibition and activity of DAOCS mutants at high penicillin G concentration provide important insight to help select proper mutants for industrial application.
Catalysis
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Intramolecular Transferases
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genetics
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Mutation
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Penicillin G
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pharmacology
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Penicillin-Binding Proteins
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genetics
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Streptomyces
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enzymology
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genetics
8.Heterologous expression and substrate specificity of ketoreductase domain in bacillaene polyketide synthase.
Xiaohui SUN ; Chengchuan CHE ; Junjie JI ; Jianting ZHENG ; Ge YANG
Chinese Journal of Biotechnology 2015;31(9):1355-1362
The ketoreductase (KR) domain in the first extending module of the polyketide synthase (PKS) catalyzes the reductions of both an α-keto group and a β-keto group in the biosynthesis of bacillaene, suggesting the intrinsic substrate promiscuity. In order to further investigate the substrate specificity, the KR domain (BacKR1) was heterologously overexpressed in Escherichia coli. In vitro enzymatic analysis showed that only one of the four diastereomers was formed in the reduction of the racemic (±)-2-methyl-3-oxopentanoyl-N-acetylcysteamine thioester catalyzed by BacKR1. In addition, BacKR1 was revealed to catalyze the reductions of cyclohexanone and p-chloroacetophenone, indicating the potential of KR domians of PKSs as biocatalysts.
Bacterial Proteins
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genetics
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metabolism
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Catalysis
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Cyclohexanones
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metabolism
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Escherichia coli
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enzymology
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Polyketide Synthases
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genetics
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metabolism
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Protein Structure, Tertiary
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Substrate Specificity
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omega-Chloroacetophenone
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metabolism
9.Comparative study on intermediate-term clinical effects of different methods in arthroscopy assisted anterior cruciate ligament reconstruction
Yao JIANG ; Jinzhong ZHAO ; Hao SHEN ; Junjie SHAO ; Ji SHEN
Chinese Journal of Trauma 1991;0(02):-
Objective To study the 2-4 years intermediate-term clinical effects of bone-patellar tendon-bone (B-PT-B) autograft plus interface screw method and quadruple semitendinosus tendon autograft plus endobutton plate fixation method in anterior cruciate ligament (ACL) reconstruction after ACL injury and to estimate the correlation of various implants and fixation techniques with clinical effect. Methods A retrospective analysis was done on 104 cases with ACL injury as well as detailed clinical data from October 1998 to October 2000. ACL reconstruction with bone-patellar tendon-bone autograft (PT group) and kurosaka interface screw fixation were performed in 55 cases and quadruple semitendinosus tendon autograft and endobutton plate fixation (ST group) in 49 cases. The average follow up was 38.5 months in the PT group and 35.7 months in the ST group. We compared the intermediate myodynamic recovery, ROM, lachman test, pivot shift test, IKDC score, lysholm score, Werner score and X-ray view of tunnul enlargement between two groups. Results In the PT group, the circumference of thigh was (1.097?0.079) cm, shorter than that of normal side, and (0.896?0.126) cm in the ST group. The cases accounting for 82% (45/55) in the PT group and 88% (43/49) in the ST group had extension limitation less than 3?. Knee flexion limitation of 0?-5? accounted for 87% (48/55) in the PT group and 86% (42/49) in the ST group. Bilateral comparison of Lachman test less than 5 mm accounted for 93% (51/55) in the PT group and 96% (47/49) in the ST group. Negative rate of pivot shift test was 89% (47/55) in the PT group and 92% (45/49) in ST group. Bilateral comparison of AP knee laxity with 20Ib KT-1000 in laxity with less than 3 mm accounted for 53% (29/55) in the PT group and 57% (28/49) in the ST group. Lyoholm score in the PT group was increased from preoperative 69.2?6.4 to postoperative 86.7?4.3 and that in the ST group from 68.3?7.5 to 88.4?8.6. Knee motion standard of IKDC score in the PT group was 78%(43/55), which was found normal or nearly normal, and 82%(40/49) in the ST group. Patellar femoral pain Werner score was 43.0 ?6.1 in the PT group while 45.0?6.3 in the ST group. The X-ray showed that tunnel enlargement phenomena was 53% (29/55) in the PT group while 43% (21/49) in the ST group. Conclusions From 2-4 years intermediate-term follow up, there is no significant difference in ACL reconstruction using patellar tendon autograft plus interface screw method and quadruple semitendinosus tendon plus endobutton plate fixation method. Bone grafting in the donor site can reduce the incidence of anterior knee pain. Twisting technique is beneficial to overcome of the postoperative laxity caused by viscular elasticity of quadruple semitendinosus.
10.TRAIL and its receptor expression in activated lymphocytes
Junjie JI ; Lixin WEI ; Xiaodong LI ; Mengchao WU ; Yajun GUO ;
Academic Journal of Second Military Medical University 2000;0(07):-
Objective: To investigate the expression difference of TRAIL and its receptors between resting and activated peripheral blood lymphocytes, and to study their role in cellular growth and proliferation. Methods: Expression of TRAIL and its receptors were detected by RT PCR and DNA sequencing. Results: Neither the expression of TRAIL nor its receptors were detected in resting peripheral blood lymphocyte. However, both expression were found in activated peripheral blood lymphocytes of various degree. Conclusion: TRAIL and its receptors may play a role in regulation and control of the activated lymphocytes in immune reaction.