1.Establishment of the test method of the Aβ42 in the serum of the alzheimer disease patient and its clinical significance
Huacheng WANG ; Jingang LUO ; Xuejun LIU ; Mengxin YANG ; Ying WANG ; Ying XU ; Chaohui DUAN
Chinese Journal of Laboratory Medicine 2012;35(1):42-46
ObjectiveTo establish a enzyme-linked immunosorbent assay (ELISA) method for detecting the β-amyloid peptide 42 ( Aβ42 ) and explore its clinical meaning for diagnosis and treatment in the early stages of the alzheimer disease ( AD).Methods Using the Aβ42 single chain variable fragment constructed by phage antibody library display system as coat antibody,associated with the Aβ42 polyclonal antibody acquired by Aβ42 immunized rabbit and HRP labeled goat anti rabbit IgG to establish ELISA method for detecting the Aβ42 in peripheral blood.The method was used it to test the Aβ42 in 120 vascular dementia VD) or cerebral vessel infarction patients and 120 AD patients and 120 controls.The methodology performance were evaluated.ResultsThe inter and intra coefficient of variable (CV) of this self-established ELISA method was 3.6% and 3.5%,6.8% and 7.1% respectively.The recovery rate was 97.2% -103.1%.The linear range was 0.050 - 2 μg,/L.Its reactivity decreased < 12% when it was put in both 37 ℃ for 6 days and 4 ℃ for 6 months.Compared with the Belgium INNOTEST reagent by testing 90 samples simultaneously,the results of self-established method was (0.207 ± 0.039 ) μg/L,the results of INNOTEST was (0.206± 0.038 ) μg/L; the regression equation was Y =1.011X - 0.003,R2 =0.979,P <0.01.The Aβ42 in blood of AD group was (0.247 ± 0.032 ) μg/L,VD or cerebral vessel infarction group was (0.173 ±0.028) μg/L,control group was (0.172 ±0.032) μg/L.The Aβ42 in AD group was higher than that in the VD or cerebral vessel infarction group and control group (q =18.867,18.907respectively,P < 0.01 ).The cut off value was 0.212 μg/L decided by the receiver operating characteristic (ROC) curve.The reference interval was 0 -0.212 μg/L.The sensitivity of this ELISA method was 86.7%(104/120) and specificity was 90.8% (218/240).ConclusionsThe ELISA method for detecting Aβ42 in peripheral blood established by the study is sensitive and specific and has good precision and stability.It could provide a new effective criterion and support for the early diagnosis and treatment of the AD patients.
2.Correlation between CYP2D6 polymorphism and risperidone metabolism in schizophrenic patients
Ting ZHANG ; Qingmin RAO ; Ying XU ; Mengxin YANG ; Pingying QIU ; Yulong LIN
Chinese Journal of Laboratory Medicine 2019;42(4):306-311
Objective To investigate the effect of CYP2D6 gene polymorphism on risperidone (RISP) metabolism in schizophrenic patients. Methods CYP2D6 allele polymorphisms including*10,*4,*41 and *2 was detected by real-time fluorescent PCR in 120 schizophrenic patients who have taken risperidone continually. Alleles without SNP mutations were classified as wild-type (WT). At the same time, serum risperidone and 9-hydroxyrisperidone concentration of all patients were detected by mass spectrometric analysis. Some samples were selected for DNA sequencing of CYP2D6*10, which is the most common CYP2D6 allele in Oriental population. The 120 patients were divided into three groups according to their allele variants. Group 1 was defined as carriers of two functional alleles, group 2 was defined as carriers of one defective allele, group 3 was defined as carriers of two defective alleles. Genotype distributions, alleles frequencies, RISP, 9-oh-RISP, RISP+9-OH-RISP, 9-oh-RISP/RISP among three groups were calculated.Results Group 1 amount to 23 cases including 13 cases of WT/WT, 7 cases of*2/*2, 3 cases of WT/*2. Group 2 amount to 51 cases, including 38 cases of WT/*10, 8 cases of *2/*10, 1 case of *2/*41, 4 cases of WT/*41. Group 3 amount to 46 cases, including 44 cases of *10/*10, 2 cases of *10/*41. The*4 allele was not detected. The allele frequency of WT, *2, *10 and *41 was 29.6%, 10.8%, 56.7% and 2.9%, respectively. The 9-hydroxyrisperidone/risperidone-ratio of three groups were 15.24±5.77, 11.06±4.56 and 2.39 ± 1.06, respectively. There was a significant difference in 9-hydroxyrisperidone/risperidone-ratio between Group 3 and the first two groups (P<0.001). Conclusions The frequency of *10 allele was the highest among the subjects. The frequency of WT and*2 allele was over 95%in the population. Individuals carrying one defective allele of CYP2D6 will decrease the rate of risperidone metabolism slightly, while individuals carrying two defective alleles of CYP2D6 may decrease the rate of risperidone metabolism significantly.
3.Correlation between personality traits and clinical symptoms in patients with anxious depression
Zhaosong CHU ; Xin WANG ; Mengxin HE ; Yuqi CHENG ; Xiufeng XU ; Zonglin SHEN
Chinese Journal of Behavioral Medicine and Brain Science 2022;31(1):31-36
Objective:To investigate the personality traits of patients with anxious depression and the relationship between personality traits and clinical symptoms.Methods:From December 2011 to October 2014, 177 first-episode untreated patients with depression from the psychiatric department of the First Affiliated Hospital of Kunming Medical University and 185 healthy controls(HC group) recruited by the community were included.All patients were divided into anxious depression group ( n=92) and non-anxious depression group ( n=85) according to whether the anxiety/somatization factor score ≥7.The simplified version of Neuroticism Extraversion Openness Five-Factor Inventory (NEO-FFI) and the Hamilton depression scale-17 (HAMD-17) were used to assess all the subjects.Statistical analyses were conducted in SPSS 21.0.Analysis of covariance was used to compare the differences of the scores on personality dimensions among the three groups.The relationship between personality dimensions and anxious depression was confirmed by Logistic regression, linear regression analysis and generalized linear models. Results:The differences of the scores on the four dimensions of neuroticism ( F=108.863, P<0.01), extraversion ( F=86.357, P<0.01), agreeableness ( F=50.615, P<0.01), and conscientiousness ( F=24.730, P<0.01) among the three groups were statistically significant.Further pairwise comparision showed, the score of neuroticisms was higher in the anxious depression group(43.05±8.92) and non-anxious depression group(39.85±7.21) than that in the HC group (30.16±6.25)( P<0.01, Bonferroni corrected). The scores of extroversion (31.22±6.33, 32.61±6.83), agreeableness (38.66±5.80, 39.46±6.19) and conscientiousness (39.75±6.89, 38.85±7.26) were lower in the anxious depression group and non-anxious depression group than those in the HC group (40.29±5.37, 44.79±4.68, 44.09±5.66, all P<0.01, Bonferroni corrected). The score of neuroticisms in anxious depression group was higher than that in non-anxious depression group, and the difference was statistically significant ( P<0.01, Bonferroni corrected). Logistic regression analysis with age, gender and years of education controlled showed that the score of neuroticism ( B=0.082, OR=1.085, 95% CI=1.020-1.154, P=0.009) and conscientiousness ( B=0.060, OR=1.062, 95% CI=1.006-1.120, P=0.028) were risk factors for anxiety symptoms in patients with depression.Linear regression analysis showed that the scores on neuroticism had positive predictive effects on the anxiety/somatization factor score ( B=0.055, 95% CI=0.021-0.089, P=0.002) and cognitive impairment factor score ( B=0.074, 95% CI=0.023-0.125, P=0.005) in the anxious depression group. Conclusion:Compared to non-anxious depression, patients with anxious depression show higher level of neuroticism, and the level of neuroticism can positively predict the symptoms of anxiety and cognitive impairment.The high level of neuroticism and conscientiousness may be risk factors for the occurrence of anxiety symptoms in patients with depressed.
4.Preparation and use of the 225Ac- 213Bi generator
Junyi CHEN ; Yu LIU ; Mengxin XU ; Zhibo LIU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2022;42(6):330-334
Objective:To develop radiopharmaceuticals for targeted alpha therapy (TAT), a simple and useful 225Ac- 213Bi generator was developed to provide a short half-life α-emitter 213Bi. Methods:DIPEX resin was used as the stationary phase of the generator, a Sep-Pak Alumina N Plus Light Cartridge column was utilized as the column to construct the generator, and hydrochloric acid was used as the eluent. The γ spectrometer was used to determine the purity of the eluted 213Bi and verify the method to measure the radioactivity of 225Ac and 213Bi by using radioisotope calibrators. The elution curves of the generator at various settings were analyzed to find the best elute and storage conditions. Results:Using commercial 225Ac, 100 mg DIPEX resin and a Sep-Pak Alumina N Plus Light Cartridge column, a 225Ac- 213Bi generator with a column volume of 0.35 ml was constructed, which could be eluted 213Bi with a syringe drenched with hydrochloric acid. The produced 213Bi was pure, and the measured half-life was (45.75±0.12) min, which was consistent with the theoretical value (45.6 min). CRC ? calibrators with setting numbers of #086 and #018 could measure the activity of 225Ac and 213Bi with the average errors of -2.6% and + 0.2% respectively. The eluent for the 225Ac- 213Bi generator was 2 mol/L hydrochloric acid, of which 320 μl was required only to get 90% elution yield. The generator was recommended to be stored directly in the eluent to avoid drying. Conclusion:The 225Ac- 213Bi generator can be constructed to obtain 213Bi for TAT.
5.Identification and verification of key salt-tolerant amino acid sites of banana MaNHX5.
Ya XU ; Mengxin TENG ; Fei QIAO ; Xingyu JIANG ; Yang ZHOU ; Xinguo LI
Chinese Journal of Biotechnology 2022;38(5):1946-1952
In order to improve the salt tolerance of banana NHX genes, we cloned a MaNHX5 gene from Musa acuminata L. AAA group and predicted the key salt-tolerant amino acid sites and mutant protein structure changes of MaNHX5 by using bioinformatics tools. The 276-position serine (S) of MaNHX5 protein was successfully mutated to aspartic acid (D) by site-directed mutagenesis, and the AXT3 salt-sensitive mutant yeast was used for a functional complementation test. The results showed that after the mutated MaNHX5 gene was transferred to AXT3 salt-sensitive mutant yeast, the salt tolerance of the mutant yeast was significantly improved under 200 mmol/L NaCl treatment. It is hypothesized that Ser276 of MaNHX5 protein plays an important role in the transport of Na+ across the tonoplast.
Amino Acids/metabolism*
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Gene Expression Regulation, Plant
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Musa/metabolism*
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Plant Proteins/metabolism*
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Plants, Genetically Modified
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Saccharomyces cerevisiae/metabolism*