3.Elemental Fractionation Studies of Biological Samples UsingLaser Ablation Inductively Coupled Plasma Mass Spectrometry
Qing LI ; Guoxia ZHANG ; Yirui CHEN ; Zheng WANG ; Chuanxian DING
Chinese Journal of Analytical Chemistry 2017;45(6):868-873
The ablated aerosols of biological matrix sample were studied using 213 nm nanosecond laser ablation system.The stable signal intensity and high sensitivity were obtained when the laser energy was 25%, the spot size was 200 μm, the scan rate was 20 μm/s, the frequency was 20 Hz and the carrier gas was 700 mL He + 700 mL Ar.Relative fractionation index of 56 elements were investigated and 31P as the internal standard element was selected under the optimized laser ablation conditions.The results showed that particle size of the biological sample was 3 μm, which was larger compared with NIST 610 sample.Element fractionation in biological sample was smaller than in glass sample, and relative fractionation index of most elements attained 1.0 ± 0.1.Element fractionation mechanism of biological sample was discussed.The possible reason why the relative fractionation index in biological sample with large particle size did not significantly increase compared to the glass sample is that the 3-μm particles entered into ICP can be atomized.On the other hand, enrichment effect for large ablation particles was relatively small.Further study of the influence factors of fractionation effect indicated that, the fractionation effect had relations with laser ablation energy, laser frequency and scan rate, negatively relation with the oxide boiling point, and positively relation with oxide bond energy and ionization energy.
4.Impact of regulatory T-lymphocytes on therapeutic efficacy of human acute brucellosis
Feihuan HU ; Guoxia ZHANG ; Yanjun TIAN ; Lijie ZHANG ; Bailing LIU ; Wenjing WANG ; Chengyao LI
Chinese Journal of Infectious Diseases 2017;35(3):129-133
Objective To investigate the humoral and cellular immune responses in patients with acute brucellosis, and evaluate dynamic changes of regulatory T-lymphocytes (Foxp3+ Treg) in the peripheral blood of patients during treatment, in order to clarify the relationship between immunosuppression and the therapeutic effect in human brucellosis.Methods Sixty-five patients with brucellosis hospitalized at the Third Department of Infectious Diseases, Heilongjiang Agriculture and Reclamation Bureau General Hospital between July 2015 and November 2015 were included.Twenty-eight patients were treated with conventional therapy (group A: patients received 3 courses of treatment.Each lasted for 20 days with one-week interval), and 37 patients were treated with conventional therapy in combination with immunopotentiator (group B).Thirty healthy volunteers were enrolled as the controlled group.The ratio of CD3+CD4+ Foxp3+ Treg cells in the peripheral blood of brucellosis patients were measured by flow cytometry (FCM) at the end of each course of treatment.Data in accordance with normal distribution were described as mean±standard deviation.Comparison between two groups was done by two sample t test.Comparison among multiple groups was performed by analysis of variance and SNK test.Data that did not fit the normal distribution were analyzed by multiple-sample nonparametric test.Results After the first (20 d), second (50 d) and third course of treatment (80 d), the ratios of Foxp3+Treg in the peripheral blood of 65 acute brucellosis patients were 2.83%, 3.77% and 4.03%, respectively, which were all significantly higher than control group (1.69%;t=5.97, 9.05 and 5.66, respectively, all P<0.01).At the end of the first course of treatment, the ratios of Foxp3+Treg in group A and B showed no statistically difference (t=0.33, P>0.05), while those were both higher than control group (t=7.09 and 4.94, respectively;both P<0.01).At the end of the second course, the ratio of Foxp3+ Treg in group B was higher than group A (t=2.22, P<0.01), and both of them were higher than control group (t=10.79 and 7.25, respectively;both P<0.01).At the end of treatment, Foxp3+ Treg in group A was also significantly higher than the other two groups (t=6.02 and 6.45, respectively;both P<0.01).Conclusions In patients with acute brucellosis treated with the standard antibiosis treatment in combination with immunopotentiator, the ratio of Foxp3+Tregs significantly increases and maintains at a high level, which suggests that extra immunopotentiator may be not helpful for the treatment of brucellosis at the very early stage.
5.Effect of psychological stress factors in the attack of systemic sclerosis
Yue CHEN ; Jizong HUANG ; Qiaoyun ZHANG ; Feixue ZHOU ; Xiaozhi QIAO ; Yu QIANG ; Guoxia GUO
Chinese Journal of Tissue Engineering Research 2006;10(6):162-164
BACKGROUND: The real mechanism of systemic sclerosis is still not clear, it is found clinically that there are psychological stress factors of different forms before the attack.OBJECTIVE: To deeply investigate the psychosocial factors in patients with systemic sclerosis, and primarily analyze the role of psychological stress factors by comparing with national norms and controls as well as combining with clinical immunological test.DESIGN: A controlled observation.SETTING: The experiments were carried out in the Department of Geriatrics, the First Hospital Affiliated to Medical College, Zhejiang University and the Department of Surgery, Hangzhou Tongji Hospital.PARTICIPANTS: Between December 2002 and September 2005. 26 patients with systemic sclerosis (systemic sclerosis group), who received thoracic duct lymph drainage therapy, and 30 inpatients with chronic gastritis(control group) were selected from the same disease area of the Department of Internal Medicine, the First Hospital of Medical College, Zhejiang University.METHODS: All the patients filled the general information inventory, the contents included were age, gender, main symptoms, disease course, special examination, diagnosis and drug therapy. The psychosocial scale was used to evaluate the psychosocial factors. Life event scale (LES) was used to assess the life events stress, including profession, learning, marriage and love, family and child, economics, justice, interpersonal relationship and other common life events. The simplified coping style questionnaire (SCSQ)was applied to assess the coping styles, including 8 main components: confrontation, indifference, self-control, seeking help, self-blame, escape, planning and reassessment, and then the habitual coping styles were divided into negative ones and positive ones. The Eysenck personality questionnaire (EPQ) was used to evaluate the personality characters, it consisted of4 subscales: extraversion-introversion scale, neuroticism scale, psychoticism scale, lie and cover up scale. The fasting blood samples (3 mL) were drawn from the patients to detect serum levels of immunoglobulin G (IgG), immunoglobulin A (IgA) immunoglobulin M (IgM) and complement C3 with immunoturbidimetry, and the correlations between IgG and other factors were analyzed.MAIN OUTCOME MEASURES: The evaluative results of psychosocial scale and results of clinical immunological detection were mainly observed.RESULTS: All the 56 patients finished the scale survey and immunological detection, and all were involved in the analysis of results. The total number of life events, number of negative events and LEU value of negative events in LES were all significantly greater in the systemic sclerosis group than in the control group. For the coping style, the dimension of positive coping was less but that of negative coping was more in the systemic sclerosis group than in the control group. For EPQ, the scores of extraversion-introversion were lower but the scores of neuroticism in both males and females in the systemic sclerosis group were higher than in the control group and norms. For the immunological detection, the levels of lgG, lgA and lgM were all higher in the systemic sclerosis group than in the control group, but C3 level had insignificant difference between the two groups.IgG had negative correlations with the number of negative events, dimension of negative coping and the score of extraversion-introversion in EPQ.CONCLUSION: Patients with systemic sclerosis have obvious psychological stress, negative coping style, unstable mood and abnormal humoral immune function. Psychosocial stress has influence on immunology, it is indicated that psychological stress is closely correlated with the attack of systemic sclerosis.
6.Determination of Trace Metals in Lutetium-Yttrium Orthosilicate by Solution-Cathode Glow Discharge-Atomic Emission Spectrometry
Rongyin GAI ; Zheng WANG ; Yanfeng HE ; Qing LI ; Huijun ZOU ; Guoxia ZHANG
Chinese Journal of Analytical Chemistry 2014;(11):1617-1622
TheLutetium-YttriumOrthosilicate(LYSO)isakindofscintillatingcrystalmaterialwiththebest comprehensive properties. In this work, the trace elements Ca, Fe, K, Li, Mg and Na in LYSO were determined by solution-cathode glow discharge-atomic emission spectrometry ( SCGS-AES ) . The optimal conditions included 0. 1 mol/L HNO3 sample solutions and operation at a voltage at 1080 V with a flow rate of 2. 0 mL/min. The LYSO matrix concentration tolerance of the SCGD source was determined to be 10 g/L. Sample solutions dissolving from several LYSO samples with HF, HNO3 and HClO4 were examined by SCGD-AES. For LYSO samples, the values obtained by SCGD-AES agree well with those obtained by axial inductively coupled plasma-atomic emission spectroscopy ( ICP-AES ) and inductively coupled plasma-mass spectroscopy (ICP-MS). The emissions of Lu and Y were not observed, so that the determination of trace elements in LYSO matrices could be conducted with little interference. The detection limits of Ca, Fe, K, Li, Mg and Na in LYSO were 1. 0, 3. 0, 0. 02, 0. 01, 0. 02 and 1. 0 mg/kg, respectively.
7.Establishment and application of serum antibodies detection methods of diphtheria and tetanus
Yajun TAN ; Deju XIA ; Huajie ZHANG ; Guoxia DONG ; Zhe CHAO ; Lin TIAN ; Qiming HOU ; Xiao MA
International Journal of Laboratory Medicine 2017;38(16):2237-2239,2242
Objective To establish the enzyme-linked immunosorbent assay (ELISA) methods for the quantitative determination of IgG antibodies against diphtheria (DT) and tetanus (TT).MethodsPurified diphtheria toxiod and tetanus toxoid were respectively used as the coating antigens,the human-derived serum antibody standard substance of DT and TT served as the standard substance.The dose-response curves of the tested samples and standard substance were fitted.Then the two quantitative ELISA methods for determining the antibody to DT (Anti-DT) and antibody to TT (Anti-TT) were established with the parallel lines method.Then the methodological verification and application study were conducted.Results The validation results of the two quantitative ELISA measurement methods were in accordance with the regulations.The quantity limit of ELISA method for quantitative detection of Anti-DT demonstrated to be 0.084 mIU/mL,its average recovery rate was 97.6%.The intra-assay coefficient of variation(CV) and inter-assay CV of this Anti-DT assay were ≤ 3.40% and ≤5.05%,respectively.The quantity limit of ELISA method for quantitative detection of Anti-TT demonstrated to be 0.175 mIU/mL,its average recovery rate was 97.5%.The intra-assay CV and inter-assay CV of this Anti-TT assay were ≤ 2.42% and ≤5.58%,respectively.These two methods were applied for the immunogenicity evaluation after infantile basic immunization by diphtheria and tetanus vaccines.Conclusion The two established quantitative ELISA methods demonstrate high accuracy and good reproducibility,which are suitable for the ordinary laboratory to carry out the work and can be used in the serological effect evaluation after diphtheria and tetanus vaccine immunization and epidemiological study of diphtheria and tetanus disease.
8.Content Determination of Four Flavonoids in Hedysari Radix in Gansu Province Based on Quantitative Analysis of Multi-components by Single Marker
Xiujuan YANG ; Jing SHAO ; Zhijun YANG ; Guoxia WU ; Yanmei NING ; Jinbao ZHANG ; Shengrui HEI
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(8):66-69
Objective To establish a method for simultaneous determination for the contents of four flavones (ononin, calycosin, genistein and formononetin) of Hedysari Radix in Gansu Province with quantitative analysis of multi-components by single-marker (QAMS); To prove its feasibility and accuracy. Methods Calycosin was taken as internal standard substance. Relative correction factors (RCF) of ononin, genistein and formononetin to calycosin were established. The contents of ononin, calycosin, genistein and formononetin were determined to realize QAMS. Results RCF was with good repeatability. The results of QAMS were consistent with the results of the external standard method. Conclusion The method that determines the contents of ononin, genistein and formononetin with calycosin as internal standard substance, can be used for quantitative analysis of Hedysari Radix.
9.Effects of Treg cells on treatment of chronic brucellosis patients
Heng YANG ; Feihuan HU ; Guoxia ZHANG ; Yanjun TIAN ; Lijie ZHANG ; Bailing LIU ; Wenjing WANG ; Chengyao LI
Chinese Journal of Endemiology 2019;38(4):273-277
Objective To investigate the percentage of regulatory T cells (Treg) in peripheral blood lymphocytes of patients with chronic brucellosis and the percentage change before and after treatment of different regimens,and to analyze the influence of Treg cell-induced immunosuppression on the therapeutic effect of chronic stage brucellosis.Methods Using case-control study,35 patients with chronic brucellosis who were hospitalized in Heilongjiang General Hospital of Agriculture Bureau [28 males,7 females,aged (45.37 ± 20.16) years old] were selected as case group.According to the treatment regimen,they were divided into standard treatment group (15 cases) and immune enhancer group (20 cases),the treatment was 20 d;30 cases of in-hospital health examinations were selected [16 males and 14 females,aged (35.53 ± 11.38) years old] as control group.Peripheral blood sample of the subject was collected before and after the treatment,the Treg cells as a percentage in peripheral blood lymphocytes were detected by flow cytometry.And the percentage change of Treg cells of brucellosis patients who underwent different treatment regimens was analyzed.Results Before treatment,the percentage of Treg cells in peripheral blood lymphocytes of the control group,the standard therapy and the immune enhancer groups [(1.69 ± 0.38)%,(3.12 ± 0.86)%,(3.05 ± 1.07)%] was significantly different (F =25.89,P < 0.05);compared with the control group,the percentage of Treg cells in the peripheral blood lymphocytes of the standard treatment group and the immune enhancer group increased (P < 0.05);there was no significant difference between the standard treatment group and the immune enhancer group (P > 0.05).After treatment,the percentage of Treg cells in peripheral blood lymphocytes of the control group,the standard therapy and the immune enhancer groups [(1.69 ± 0.38)%,(3.06 ± 0.76)%,(2.85 ± 0.89)%] was significantly different (F =30.84,P < 0.05);compared with the control group,the percentage of Treg cells in the peripheral blood lymphocytes of the standard treatment group and the immune enhancer group increased (P < 0.05);there was no significant difference between the standard treatment group and the immune enhancer group (P > 0.05),and compared with the same group before the treatment,respectively,the differences were not statistically significant (P > 0.05).Conclusions The percentages of Treg cells in peripheral blood lymphocytes of the chronic brucellosis patient are not significantly changed before and after different treatment regimens.It suggests that the immunesuppression induced by Treg cells may be one of the reasons why the host organism cannot effectively remove residual Brucella in the body,which leads to chronic infection.
10.Clinical phenotype and variantal analysis of a pedigree affected with hereditary coagulation factor V deficiency.
Fengyu CHE ; Wendi HUANG ; Ying YANG ; Guoxia WANG ; Liyu ZHANG ; Ruobing LIANG ; Jiangang ZHAO
Chinese Journal of Medical Genetics 2020;37(4):427-430
OBJECTIVE:
To explore the molecular basis for a pedigree affected with coagulation factor V (FV) deficiency.
METHODS:
Clinical data of the patient and his family members was analyzed. Targeted capture and next-generation sequencing (NGS) and Sanger sequencing were carried out to detect potential variant of the FV gene.
RESULTS:
The patient presented with jaundice and prolonged prothrombin time (PT) and activated partial thromboplastic time (APTT). V factor activity measured only 0.1% of the normal level, though the patient had no sign of bleeding. A paternal heterozygous variant c.653T>C (p.F218S) and a maternal heterozygous variant c.3642_3643del (p.P1215Rfs*175) were identified in the FV gene of the patient. His elder brother was a heterozygous carrier of the c.653T>C (p.F218S) variant. c.653T>C(p.F218S) was a known pathogenic variant, while the c.3642_3643del (p.P1215Rfs*175) variant was unreported previously.
CONCLUSION
Mutations of the FV gene probably underlie the hereditary coagulation factor V deficiency in this patient. NGS combined with Sanger sequencing has detected potential variant with efficiency and provided a reliable basis for clinical and prenatal diagnosis for this family.
Aged
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Factor V
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Factor V Deficiency
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genetics
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Genetic Variation
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Heterozygote
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Humans
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Male
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Mutation
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Pedigree
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Phenotype