1.Prognostic significance of increased ring sideroblast in myelodysplastic syndrome
Sujun GE ; Mianyang LI ; Huiyuan KANG ; Lilin GU ; Yuling PAN ; Gaixia LIU ; Wencan JIANG ; Shuang LIANG ; Chengbin WANG
Chinese Journal of Laboratory Medicine 2017;40(2):126-132
Objective This study is aimed to investigate the prognostic significance of ring sideroblast ( RS) in MDS( Myelodysplastic Sydrome ) and evaluate the correlation of RS and other prognostic index.Methods A total of 198 patients with MDS between March 2009 and December 2015 in Chinese PLA′s Gerneral hospital were chosen for this study .Based on the ratio of RS in nucleated red blood cell , patients were first separated into myelodysplastic syndrome without ring sideroblast (MDS RS-) group, RS≥15%, and myelodysplastic syndrome with ring sideroblast ( MDS RS +) group, RS <15%. Then, according to the proportion of blasts in bone marrow nucleated cells above 5%or below, patients were further divided into myelodysplastic syndrome with low blasts without ring sideroblast ( MDS-LB RS-) group, myelodysplastic syndrome with low blasts and ring sideroblast ( MDS-LB RS+) group, refractory anemia with excess blast without ring sideroblast ( RAEB RS-) group and refractory anemia with excess blast and ring sideroblast ( RAEB RS+) groupe.All patients had completed the morphological , genetics , molecular biology examination at dignosis, and followed up by phone.The results of the overall survival (OS) analysis have been presented in a Kaplan-Meier curve and cox regression model .Last, according to the percentage of RS in nucleated red blood cell , patients were separated into RS <5%groupe, 5%-15%group, 15%-40%group, RS≥40%group, and analyse their survival prognosis by statistical methods .Results Comparing to MDS RS-group, the morbidity age, WBC and PLT count were significantly higher [61 ±1.91 vs 52 ±1.37, t=-3.555, P<0.01, 3.82(0.47-323)vs 2.6(0.6-59.7), z=-4.014, P<0.01;139.5(7-608) vs 60(3-724), z =-3.988, P<0.01], bone marrow eythroid hyperplasia and gigantocyte were more obvious in MDS RS+group[χ2 =11.032, P<0.01, χ2 =5.165, P<0.05]; the percentage of GATA1 gene and abnormal rate of poor prognosis gene ( MLL, NRAS, WT1 ) , either mutation or high gene expression , were higher in MDS-LB RS+group than that in MDS-LB RS-( P<0.05 ); Contrasting with RAEB RS-group, the karyotype is worse in RAEB RS +group[χ2 =4.966, P<0.05];Comparing to 15%-40%group, the OS were poorer in RS≥40%;MDS RS+patients were more prone to adverse prognosis than MDS RS-patients.Conclusion Compared to MDS RS-group, MDS RS +patients had worse prognosis;RS maybe correlate to morbidity age , eythroid dysplasia and gene abnormality in affecting the survival prognosis of MDS.
2.Application of element-tagged immunoassay combined with inductively coupled plasma mass spectrometry in HCG detection
Danna PU ; Wencan JIANG ; Xinyu WEN ; Guanghong GUO ; Ping LI ; Chengbin WANG
Chinese Journal of Laboratory Medicine 2020;43(3):284-290
Objective:To establish an inductively coupled plasma mass spectrometry (ICP-MS) based immunoassay method for the quantitative detection of human chorionic gonadotropin (HCG), and evaluate the clinical applicability of this method.Methods:Sm was selected as element tags, and the HCG quantitative detection system was established by double antibody sandwich method. The dosage of biotinylated antibody and reaction time were optimized. According to EP documents of Clinical and Laboratory Standards Institute (CLSI) and related standards, the analytical performance was evaluated after the establishment of the assay, including the limit of blank (LOB), linearity, precision, recovery, cross reactivity and interference test. And 88 clinical samples were measured using the new method compared to the electrochemical immunoassay (ECLIA) method.Results:Total process completed within 30 min after optimization, and the optimal biotinylated antibody dosage was 0.5 μl. The LOB was 0.29 mIU/ml. The linearity was good within the range of 1.16-8 365.62 mIU/ml with the linear correlation coefficient greater than 0.995 ( R2=0.998 0), the recovery was 97.53%-102.01%. Both intra-and inter-assay coefficients of variation of the high-value sample and the low-value sample were less than 10%. And there was no significant cross-reaction with Luteinizing hormone (LH), follicular stimulating hormone (FSH), and thyroid-stimulating hormone (TSH). The interference bias caused by different concentrations of interference substances was less than 10%. When compared with the ECLIA method for clinical sample detection, the proposed method showed a significant correlation( R2=0.960 0). Conclusion:The proposed ICP-MS base immunoassay for HCG detection has good accuracy, high sensitivity and specificity, and the results of analytical performance verification meet the clinical requirements, which provides experimental basis for the clinical application of this method.
3.Intergration of co-immunoprecipitation technology and mass spectrometric analysis for sperm membrane protein screening
Shuang LIANG ; Daijun XIANG ; Hongxia WANG ; Junlong MA ; Dandan XUE ; Peipei LIU ; Ping LIU ; Wencan JIANG ; Xiaozhou YUAN ; Xinjun LI ; Sujun GE ; Chengbin WANG
Chinese Journal of Laboratory Medicine 2017;40(4):298-302
Objective To reveal the mechanisms of immunological infertility,the method of coimmunoprecipitation(CO-IP) and liquid chromatogram mass/mass (LC-MS/MS) was used to screen sperm membrane proteins which interacting with antisperm antibodies (ASA).Methods This study was designed as a case-control.The disease group including 56 serum samples from 521 cryptogenic infertile patients were screened ASA positive by ELISA and conformed with mixed antiglobulin reaction(MAR).The controls were 31 serum samples which ASA is negative and already possessed healthy offspring.All subjects were enrolled from September 2015 to December 2015 in China PLA General Hospital.Spermatozoa samples from 48donors with normal sperm parameters were from January 2016 to April 2016 in China PLA General Hospital.The purified human sperm membrane proteins were then mixed with serum from disease group (positive for ASA) and control group (not containing ASA).The binding proteins of antisperm antibodies were enriched using CO-IP assay.The immunoprecipitates were separated on sodium dodecyl sulfate-polyactylamide gel (SDS-PAGE),then the binding proteins were cut from the gel and analyzed by LC-MS/MS after the enzymolysis.These proteins could bc idcntified as definition,biological function (s) and subccllular localization with Uniprot database.Results The serum samples from infertile persons (39 females and 17males) were screened ASA positive by ELISA and conformed with MAR.The healthy controls (17 females and 14 males) were ASA-negative in ELISA.Forty proteins that interact with ASA were obtained from the study and these could be divided into three groups:11 antigens detected by control serum samples only,14antigens recognised by both infertile patients and control sera,and 15 antigens specific for patients with ASA.These 15 proteins are Sperm Cation channcl protein 1,Sperm Cation channel protein 3,Sperm Cation channel protein 4,Sperm associated antigen 9,Apolipoprotein A-I,Dynein heavy chain 14,Cylicin-2,Izumo sperm-egg fusion protein 4,Thioredoxin domain-containing protein 2,IQ domain-containing protein H,IQ domain-containing protein F1,Spermatogenesis-associated protein 5,Spermatogenesis-associated protein 5-like protein 1,Sperm acrosome membrane-associated protein 1,E3 ubiquitin-protein ligase RNF 114.Conclusion Fifteen proteins discovered with CO-IP technology and LC-MS/MS analysis could be referred as male immunoinfertility-related antigens and they may hold the great importance in revealing the secret of immunological infertility.
4.Prevalence and factors associated with HIV testing uptake among sexual active college students in Zhuhai City
Chinese Journal of School Health 2021;42(8):1246-1250
Objective:
To understand the prevalence of HIV (human immunodeficiency virus) testing and associated factors among sexual active college students in Zhuhai City.
Methods:
From November to December, 2019, an anonymous electronic questionnaire was administered among 12 235 students in six colleges and universities in Zhuhai City by multistage sampling. A total of 1 789 college students ever had sex were selected. Pearsons Chisquare test and nonconditional Logistic regression model were applied to analyze the factors associated with uptake of HIV testing.
Results:
Among these students ever had sex, 7.55% (135/1 789) had been tested for HIV mainly through hospitals (71.85%). The main reasons for testing were regular testing (50.37%) and intending to know their infection status (23.70%). Multivariable Logistic regression showed that homosexual individuals (OR=4.62, 95%CI=1.07-19.95) and those who had heterosexual commercial sex in the past year (OR=3.77, 95%CI=1.96-7.26) were more likely to test for HIV, while female (OR=0.41, 95%CI=0.24-0.69) were less likely to test for HIV.
Conclusion
The proportion of HIV testing was low among sexual active college students in Zhuhai City. Interventions should be tailored including strengthening the HIV testing propaganda education and enhancing students awareness of HIV testing, and the influencing factors such as gender, sexual orientation and commercial sexual behavior should be taken into consideration, so as to improve the HIV testing coverage of this population.
5.Analysis of associated factors of the inconsistency for knowledge and behavior in condom use among college students in Zhuhai City
Chinese Journal of School Health 2023;44(10):1497-1500
Objective:
To examine the prevalence and factors influencing the inconsistency for knowledge and behavior in condom use among college students, so as to provide suggestions for AIDS prevention and education in universities.
Methods:
From October to December 2019, a multistage cluster sampling method was employed to collect data relating to inconsistency for knowledge and behavior in condom use and other related factors among 1 303 students from six colleges in Zhuhai, China. Chi square test and Logistic regression were performed to analyze the influencing factors and moderating effect.
Results:
The reporting rate of in consistency of knowledge and behavior in condom use among college students was 41.1%. Multivariate Logistic regression analysis showed that seeking sexual partners offline was negatively correlated with inconsistency for knowledge and behavior in condom use ( OR=0.70, 95%CI =0.51-0.95). However, condom nonuse during the first sexual experience (OR=7.11, 95%CI=5.23-9.67), smoking before sex ( OR=1.47, 95%CI =1.07-2.02), drinking before sex ( OR=1.44, 95%CI =1.09-1.91), history of intimate partner violence ( OR=1.53, 95%CI =1.13-2.07), and having multiple sexual partners ( OR=1.69, 95%CI =1.25-2.29) were positively correlated with inconsistency for knowledge and behavior in condom use ( P <0.05). The moderating effect analysis showed that condom use during the first sexual experience had a moderating effect on smoking before sex and inconsistency for knowledge and behavior in condom use ( β=0.92, P <0.05). Among students who did not use condoms during the first sexual experience, a positive correlation was observed between smoking before sex and inconsistency for knowledge and behavior in condom use ( OR= 2.76 , 95%CI=1.09-6.99, P <0.05). However, no correlation was found between smoking before sex and inconsistency for knowledge and behavior in condom use ( OR=1.32, 95%CI=0.92-1.88, P >0.05) among students who used condoms during the first sexual experience.
Conclusion
High levels of inconsistency for knowledge and behavior in condom use are found among college students in Zhuhai City. Colleges should carry out sex education activities as soon as possible, and explore new health education models to promote the transformation of their knowledge into behavior.
6.Changes in cerebrospinal fluid leukocytes and 24-hour intrathecal IgG synthesis rate in the patients with positive intrathecal IgG synthesis
Shujing LIU ; Panpan FENG ; Wencan JIANG ; Lijuan WANG ; Siwen LI ; Jin ZHOU ; Guojun ZHANG
Chinese Journal of Laboratory Medicine 2024;47(2):142-146
Objective:Laboratory evaluation of the relationship between cerebrospinal fluid and plasma indicators and intrathecal immunoglobulin G(IgG) synthesis in patients with neurological diseases, and establishment of a new diagnostic method for intrathecal IgG synthesis.Methods:This study retrospectively analyzed the content of IgG in cerebrospinal fluid samples and blood albumin in blood samples, and other test results of 410 patients with neurological diseases who visited Beijing Tiantan Hospital from 2019 to 2022. According to the results of oligoclonal bands in cerebrospinal fluid, patients were divided into intrathecal IgG synthesis group and non-intrathecal IgG synthesis group. The Mann Whitney U test was used for inter group comparison, and a bilateral test with P<0.05 indicates a statistically significant difference. Include indicators with differences between groups in logistic regression analysis, construct a predictive model, and compare it with the established quantitative formula IgG index. Results:There were significant differences in 10 indicators, including cerebrospinal fluid leukocyte count and 24-hour intrathecal IgG synthesis rate, between the intrathecal IgG synthesis group and the non-intrathecal IgG synthesis group, with P<0.05. The area under the curve (AUC) of intrathecal IgG synthesis was higher than the IgG index (AUC=0.920, 0.809, Z=31.178, P<0.001), the sensitivity was higher than the IgG index (0.825, 0.618), and the specificity was lower than the IgG index (0.876, 0.908). Conclusion:The combination of 10 indicators such as cerebrospinal fluid white blood cell count and 24-hour intrathecal IgG synthesis rate can improve the diagnostic efficacy and sensitivity of intrathecal IgG synthesis.
7.Potential biomarkers for the diagnosis and differential diagnosis of immune-mediated demyelinating diseases screened by tandem mass spectrometry technology
Yaowei DING ; Yijun SHI ; Guoge LI ; Wencan JIANG ; Guanghui ZHENG ; Guojun ZHANG
Chinese Journal of Laboratory Medicine 2022;45(1):36-44
Objective:To screen the potential biomarkers for the diagnosis and differential diagnosis of immune-mediated demyelinating diseases by tandem mass tags (TMT) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology.Methods:Twenty patients with demyelinating diseases (demyelinating group) and 10 patients with noninflammatory neurological diseases (NND group) from Beijing Tiantan Hospital affiliated to Capital Medical University from January 2020 to January 2021 were enrolled in this study. The demyelinating group included 10 patients with Guillain-Barre syndrome (GBS subgroup) and 10 patients with multiple sclerosis (MS subgroup). TMT proteomics was used to screen out the different protein expression patterns between the demyelinating group and the NND group and between the GBS subgroup and the MS subgroup (difference>2 or<0.5 and with statistical significance), and String database was used to perform gene ontology (GO) analysis and Kyoto encyclopedia of gene and genomes (KEGG) analysis on the pathways involved in the differently expressed proteins between the groups. In addition, 80 demyelinating patients (demyelinating diseases validation group) and 40 healthy subjects (healthy control group) were selected for retrospective analysis of general lipid indexes. The demyelinating diseases validation group included 40 GBS patients (GBS validation group) and 40 MS patients (MS validation group). Receiver operating characteristic (ROC) curve was obtained to evaluate the value of general lipid indexes for the diagnosis of demyelinating diseases and the differential diagnosis between GBS and MS groups.Results:A total of 362 proteins were detected by TMT proteomics. There were 101 differentially expressed proteins between the demyelinating group and the NND group, and 45 differentially expressed proteins between the GBS group and the MS group. Compared with the NND group, GO enrichment analysis showed that the top five enrichment pathways in the demyelinating group were macrophage colony stimulating factor and receptor complex, negative regulation of cholesterol input, negative regulation of very low density lipoprotein particle clearance, triglyceride-rich lipoprotein particle remodeling, and cholesterol reverse transport. Compared with MS group, the top five enriched pathways in GBS group were high-density lipoprotein particle receptor binding, negative regulation of very low density lipoprotein particle remodeling, negative regulation of cholesterol input, negative regulation of very low density lipoprotein particle clearance, and medium density lipoprotein particle. KEGG enrichment analysis results showed that differentially expressed proteins in the demyelinating group and the NND group were enriched in 8 pathways, including phosphatidylinositide 3-kinases-protein kinase B signaling pathway, complement and coagulation cascade reaction, extracellular matrix and its receptor interaction, Staphylococcus aureus infection, cholesterol metabolism, RAS signaling pathway, phagosome, and mitogen-activated protein kinase signaling pathway. Differentially expressed proteins in GBS group and MS group were enriched in 9 pathways: cholesterol metabolism, complement and coagulation cascade, platelet activation, peroxisome proliferators-activated receptors signaling pathway, vitamin digestion and absorption, novel coronavirus infection, fat digestion and absorption, axon guidance, and neutrophil extracellular trap formation pathway. The levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and apolipoprotein B (apoB) were significantly higher, while high density lipoprotein cholesterol (HDL-C) and apolipoprotein A1 (apoA1) levels were significantly lower in the demyelinating disease validation group than in the healthy control group (all P<0.05 or 0.01). Area under the curve (AUC) of TG, TC, HDL-C, LDL-C, apoA1 and apoB alone or in combination for the diagnosis of immune-mediated demyelinating diseases was 0.746, 0.643, 0.798, 0.703, 0.806, 0.708 and 0.868, respectively. The AUC of HDL-C, apoA1, LDL-C and apoB for differential diagnosis between GBS and MS was 0.692, 0.653, 0.632, 0.695 and 0.718, respectively. Conclusions:There are differences in cerebrospinal fluid proteomics between patients with immune-mediated demyelinating disease and patients with NND, GBS and MS, and the differentially expressed protein patterns mainly exist in the pathways related to lipid metabolism. Lipid related indicators may be used as biomarkers for the diagnosis and differential diagnosis of immune-mediated demyelinating disease.