1.Proteomic study for serum biomarkers in Parkinson's disease using weak cation exchange magnetic beads and MALDI-TOF-MS
Jian WANG ; Yaohua LI ; Xiaoli ZHENG ; Yanli ZHANG ; Xin LI ; Shun YU ; Xin HE ; Biao CHEN
Chinese Journal of Neurology 2009;42(2):98-101
Objective To screen for the potential protein biomarkers in serum for the diagnosis of idiopathic Parkinson's disease (PD) using proteomic fingerprint technology. Methods Proteomic fingerprint technology combining weak cation exchange (WCX) magnetic beads with MALDI-TOF-MS was used to identify and compare the serum proteins from 61 patients with idiopathic PD, 29 patients with other neurodegenerative diseases (OND) and 30 healthy blood donors. Model of biomarkers and proteomics patterns associated with PD was analyzed by Biomarker Patterns Software. The model also was validated by 40 newly recruited PD cases. Results A total of 17 discriminating M/Z peaks which were related to PD were identified ( nonparametric test, Z:-4.039--2.633, P<0.01 ). Five biomarkers with M/Z of 6121, 5234, 2961,4309 and 8170 respectively generated an excellent model of distinguishing between PD and healthy groups. The sensitivity was 98.4% and the specificity was 83.1%. Blind testing in 40 newly recruited cases demonstrated a sensitivity of 85.0% (17 of 20 PD) and a specificity of 70. 0% (14 of 20 controls). Conclusions Combination of WCX magnetic beads with MALDI-TOF-MS is a useful method in establishing proteomic patterns associated with PD. It also may be used to construct a diagnostic model with PD Biomarkers. Although this model of biomarkers fails to distinguish between PD and OND controls, it is able to differentiate PD from healthy controls.
2.Effects of heparin and low molecular weight heparin on serum level of pregnancy associated plasma protein A in patients with cerebrovascular disease
Shun WANG ; Lili WANG ; Xin ZHANG ; Chuanxin WANG ; Xuhua ZHANG ; Zhaogang DONG ; Lutao DU
Chinese Journal of Laboratory Medicine 2011;34(5):415-419
Objecfive To investigate the effects of treatment for cerebrovascular disorder patients with heparin and low molecular weight heparin(LMWH) on serum PAPP-A concentrations and provide the basis for evaluating the clinical significance of PAPP-A in the following study.Methods Forty cases with cerebrovascular disease from Qilu Hospital from November 2009 to May 2010 were collected in this study.Blood samples were taken before and after drug administration.All cases were divided into four groups according to situation of medication.Group A consisted of 10 patients who received subcutaneous LMWH anticoagulation therapy, and blood samples were collected before LMWH injection, three hours after subcutaneous LMWH anticoagulation therapy in the first day, the second day and the seventh day and 24 hours after the last injection. Group B consisted of 10 patients who did not receive LMWH therapy, and blood samples were collected immediately after admission, the first day, the second day and the seventh day after admission. Group C consisted of 10 patients with percutaneous carotid intervention who received intravenous heparin at the beginning of stenting, and blood samples were collected from the arterial sheath just before angiography and heparin administration, and at 3, 5, 15, 40 and 100 min after heparin administration. Group D consisted of 10 patients who received carotid angiography but LMWH-free therapy,and blood samples were collected from the arterial sheath just before and after angiography. Serum PAPP-A concentrations were analyzed by ELISA to evaluate the differences of intra-groups and differences at different time points of inter-groups. Results In group A, PAPP-A concentrations were time dependent and elevated gradually from 12. 36 (9. 90-14. 32) mIU/L before LMWH injection to 21.80 (23.50-19.73) mIU/L at the seventh day after injection (M=38. 72, P < 0.01 ). In group C, there was a rapid increase of PAPP-A concentration from 12. 86 ( 9. 67-14. 05 ) mIU/L to 51.56 ( 44. 20-66. 00 ) mIU/L within 5 min after intravenous heparin injection (M=46. 06, P <0. 01 ). The PAPP-A concentration of one week after LMWH administration in group A was 21.80 (23.50-19.73) mIU/L, significantly higher than that in group B [11.81 (9. 21-12. 89) mIU/L] (U<0. O01, P<0.01). The PAPP-A concentration at 15 min after heparin administration in group C was 43.70 (37.70-54. 30) mIU/L, significantly higher than that after angiography in group D [14. 18 (11.25-15. 86) mIU/L] ( U<0. 001, P <0. 01 ). The peak level of blood PAPP-A after subcutaneous LMWH injection was significantly lower than that after intravenous heparin injection. The concentrations in group A and C were 21.80 ( 23.50-19. 73 ) and 51.56 (44. 20-66. 00) mIU/L respectively, and had a significant difference ( U=0. 999, P < 0. 01 ) . Conclusions Both intravenous heparin and subcutaneous LMWH administration induce an increase in serum PAPP-A concentration. The effect of drug should be considered when PAPP-A is selected as an evaluation indicator.
3.The application of plasma soluble human leukocyte antigen in diagnosis of patients with cervical cancer and its precancerous lesion
Xin ZHANG ; Ni ZHENG ; Chuanxin WANG ; Chengbao ZHU ; Lili WANG ; Lutao DU ; Shun WANG
Chinese Journal of Laboratory Medicine 2011;34(1):61-65
Objective To explore the application value of plasma sHLA-G in diagnosis of CIN and cervical cancer. Methods The plasma sHLA-G levels were detected by ELISA in 102 cases with cervical cancer( FIGO Ⅰ stage 32 cases, Ⅱ stage 28 cases, Ⅲ stage 25 cases and Ⅳstage 17 cases; tumor size:<4 cm 63 cases and ≥4 cm 39 cases; squamous cell carcinoma 78 cases and adenocarcinoma 24 cases;cell differentiation:well 57 cases, moderate 29 cases and poor 16 cases; lymph nodes metastasis negative64 cases and positive 38 cases ), 72 cases with CIN( Ⅰ grade 21 cases, Ⅱ grade 25 cases and Ⅲ grade26 cases ) and 20 cases of healthy controls. The diagnostic value of sHLA-G and its correlations with clinical parameters were analyzed. Results The plasma levels of sHLA-G were 193.6( 151.3-287.4 ) kU/L in cervical cancer group, 48.3( 34.6-57.2 ) kU/L in CIN Ⅰ group, 91.3( 68.2-118.6 ) kU/L in CIN Ⅱ group, 106.4( 73.8-165.7 ) kU/L in CIN Ⅲ group and 45.2( 38.0-55.5 ) kU/L in health control group.The level of sHLA-G was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and healthy control group( U value of 8.832, 6.456, 4.017, 9.873, P < 0.05,respectively ). The level of sHLA-G was significantly higher in CIN Ⅱ group and CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 4.361,4.892, 5.139, 5.485, P <0.05, respectively ).The levels of SCC Ag in healthy control group, CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and cervical cancer group were 0.43( 0.38-0.69 )μg/L, 0.47( 0.35-0.72 )μg/L, 0.65( 0.53-0.81 )μg/L, 0.82( 0.54-1.03 )μg/L and 1.02( 0.62-1.87 )μg/L. The level of SCC-Ag was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group and healthy control group( U value of 7.926, 4.877, 8.132,P <0.05, respectively ). The level of SCC-Ag was significantly higher in CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 6.574, 6.763, P <0.05, respectively ). The levels of CA125 in healthy control group, CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and cervical cancer group were 14.38 ( 6.14-21.82 ) kU/L, 15.42( 6.25-23.53 ) kU/L, 21.34( 9.82-32.58 ) kU/L, 25.69( 14.47-38.71 )kU/L and 27.72( 14.29-43.87 ) kU/L. The level of CA125 was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group and healthy control group( U value of 7.564, 4.522, 7.429, P <0.05, respectively ). The level of CA125 was significantly higher in CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 5.871, 5.435, P <0.05, respectively ). ROC curve analysis showed AUC for sHLA-G was 0.828( 95% CI:0.768-0.879 ), which was high as compared with the AUC of SCC-Ag [ 0.727( 95% CI:0.658-0.788 );Z = 2.294, P < 0.05 ] and the AUC of CA125 [ 0.705( 95% CI:0.636-0.769 );Z =2.842 ,P <0.05 ]. There was no significant difference of diagnostic efficiency between SCC and CA125( Z =0.672, P > 0.05 ). When cutoff value of sHLA-G was 109.6 kU/L, the diagnostic sensitivity,specificity, positive predictive value, negative predictive value and accuracy rate were 86.3%, 76.1%,80.0%, 83.3%, and 78.4%, respectively. The levels of sHLA-G in cervical cancer patients were significantly correlated with FIGO stages and lymphoid node metastasis ( U value of 6.085, 4.451, P <0.05, respectively ), while there were no significant differences between the levels of sHLA-G and age,tumor size, histological type and cell differentiation( U value of 1.274, 1.956, 1.268, 2.719, P >0.05,respectively ). Conclusions sHLA-G can be used for the early screening of cervical cancer and its precancerous lesion. It could also be used as an index for judging progression and lymphoid node metastasis.
4.Alterations of plasma glucocerabrosidase,protein phosphastase 2A and ceramide levels in patients with ischemic stroke
Minghua ZHANG ; Yaguang SONG ; Peng WANG ; Weiwei YANG ; Xin LI ; Xuran LI ; Shun YU
Chinese Journal of Cerebrovascular Diseases 2015;(6):302-305
Objective Toanalyzethealterationsofplasmaglucocerebrosidase(GBA),protein phosphatase 2A (PP2A)and its degradation product ceramide in patients with ischemic stroke. Methods Atotalof45inpatientswithischemicstrokeattheDepartmentofNeurology,theAffiliated Hospital of Logistics College of the CAPF were enrolled from May to September 2013,and 45 age-and sex-matched healthy subjects at the Physical Examination Center collected at the same time period were used as a control group. Blood samples of the patients and healthy subjects were obtained,anticoagulated, and the plasma was separated. H50 protein chip and laser matrix-assisted laser desorption/ionization top of flymassspectrometrywereusedtotestthelevelsofplasmaceramide.Results TheplasmaGBAand PP2A activities in patients with ischemic stroke were significantly lower than those of the control group;the GBA activities of the ischemic stroke group and the control group were 2 . 4 ± 0 . 8 and 3 . 1 ± 1 . 4 U/L respectively. There was significant difference (P<0. 05);the PP2A activities of the two groups were 6. 5 ± 2. 8 and 14. 5 ± 4. 7 U/L respectively (P<0. 01). The relative level of the plasma ceramide in patients with ischemic stroke was 1. 9 ± 0. 7,and it was significantly lower than 12. 2 ± 5. 0 of the control group (P<0.01).Conclusion ThedecreasedlevelsofplasmaGBAandPP2Aactivitiesaswellasthe ceramide in patients with ischemic stroke suggested that the abnormal phosphorylation of synuclein in the blood of patients with stroke.
5.Changes of plasma phosphorylated α-synuclein level and α-synuclein phosphorylation rate in patients with ischemic stroke
Minghua ZHANG ; Weiwei YANG ; Xin LI ; Xuran LI ; Peng WANG ; Shun YU
Chinese Journal of Cerebrovascular Diseases 2014;(9):476-479
Objective Toanalyzethechangesofplasmaphosphorylatedα-synuclein(α-Syn)level andα-Synphosphorylationrateinpatientswithischemicstroke.Methods Theclinicaldataof45 patients with acute stroke admitted to the Department of Neurology,the Affiliated Hospital of Logistics University of People′s Armed Force Police from May 2013 to September 2013 were analyzed retrospectively, and the age and sex matched 45 healthy subjects were recruited as a control group at the same time. The plasma phosphorylatedα-Syn level was measured by a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA),besides,the gene-recombinated α-Syn was added into plasma,and the phosphorylated α-Syn accountingforthetotalratioofα-Synwascalculated.Results Theplasmaphosphorylatedα-Synlevelof patients with ischemic stroke was significantly higher than that of the control group (0. 0472 ± 0.0042μmol/L vs. 0. 0312 ± 0. 0043μmol/L). The plasma α-Syn phosphorylation rate of patients with ischemic stroke was higher than that of the control group (0. 1170 ± 0. 0176 vs. 0. 0364 ± 0. 0098μmol/(100μmol ·h ). Receiver operating characteristic (ROC )curve analysis showed that the specificity and sensitivity of the plasma phosphorylatedα-Syn concentration changes in determining ischemic stroke were 0. 88 and 0. 81 respectively. The area under curve (AUC)was 0. 91 and the cut-off value was 0. 060 mol/L;AUC 95%confidence interval (CI)was 0. 889 to 0. 961;the specificity and sensitivity of the plasma α-Syn phosphorylation rate changes in determining ischemic stroke were 0. 84 and 0. 81 respectively,AUC was 0.90andthecut-offvaluewas0.055mol/L;AUC95%CIwas0.898to0.971.Conclusion Theplasma phosphorylated α-Syn level and plasma α-Syn phosphorylation rate in patients with ischemic stroke were higher than those of the normal control group.
6.Hypoxic preconditioning up-regulates the activity and gene expressions of glucose transporters (GLUT1 and GLUT3) in cultured rat hippocampal neurons and astrocytes under anoxic condition
Erfu WANG ; Xin LI ; Chunsong JIA ; Guangwei LIU ; Haixia FANG ; Shun YU
Basic & Clinical Medicine 2009;29(12):1273-1276
Objective To explore the effect of hypoxic preconditioning on the activity and gene expressions of glu-cose transporters in the cultured rat hippocampal neurons and astrocytes under anoxic condition. Methods The cultured rat hippocampal neurons and astrocytes were treated for 6 days by intermittently exposing to hypoxic gas mixture (1% O_2, 10% CO_2, 89% N_2) for20 min each day. 24 h after the last hypoxic exposure, the cells were exposed to anoxic gas mixture (10% CO_2, 90% N_2) for 6 h, and the uptake rate of [~3H]-2-deoxyglucose (2-DG), the levels of glucose transporter GLUT1 and GLUT3 mRNAs and the cell survival rate were examined im-mediately after anoxic exposure. Results Neurons and astrocytes preconditioned with hypoxia showed higher 2-DG uptake rates and increased expressions of GLUT 1 mRNA in the astrocytes and GLUT 1 and GLUT 3 mRNA in the neurons. The preconditioned neurons also showed an increased tolerance to anoxia. Conclusion Hypoxic precon-ditioning up-regulates the activity and gene expressions of glucose transporters of hippocampal neurons and astro-cytes under anoxic condition.
8.Expression Levels of IL-6 mRNA in PBMNCs from Patients with IDDM,NIDDM and Normals by RT-PCR Procedure
Wei HUANG ; Deng-Shun WANG ; Xin-Yuan LI ; Wei-Zheng WU ; Gui-Cheng NI ;
Chinese Journal of Immunology 1985;0(05):-
Whether or not an abormal expression of IL-6 mRNA in PBMNCs from IDDM patientswas examined using a hihgly sensitive,specific and semiquantitative protocal,i.e.reverse tran-scription and polymerase chain reaction (RT-PCR).The relative levels of IL-6mRNA in PBM-NCs from 12 early IDDMpatients (8.20?3.85yr),29 newly diagnosed NIDDM patients(54.85?9.12yr)23 normal childrens (8.20?3.26yr) and 12 normal adults (31.92?11.22yr)weredetermined.Significantly high expresion levels of IL-6 mRNA were found in PBMNCs from pa-tients with IDDM (P
9.Meta-analysis of effect comparison between video endoscopic inguinal lymphadenectomy and open inguinal lymphadenectomy
Zixiang YAO ; Delin WANG ; Fan YANG ; Shun LIU ; Wencong LIU ; Xin ZHANG
Chongqing Medicine 2014;(12):1422-1424,1427
Objective To evaluate the effects between video endoscopic inguinal lymphadenectomy (VEIL ) and open inguinal lymphadenectomy(OIL) to provide the evidence-based basis for the selection of the clinical therapy schemes .Methods The related clinical controlled trial literature on the effective comparison of VEIL and OIL were retrieved from the databases of PubMed ,Co-chrane library ,Elsevier ,CNKI and Wanfang database .The screening was independently performed by 2 reviewers according to the including and excluding criteria .The related data were extracted and performed the meta analysis by the RevMan 5 .2 software .Re-sults A total of 4 trials were included .There were 146 cases of inguinal lymphadenectomies ,in which 61 cases were VEIL and 85 cases were OIL .The meta-analysis results showed that there were no statistical differences between the two operation modes in terms of the operative time(WMD=32 .33 ,95% CI -25 .70-90 .36 ,P=0 .27) ,intraoperative blood loss(WMD=9 .10 ,95% CI -76 .03-94 .23 ,P=0 .83) ,number of removed lymph nodes(WMD=0 .77 ,95% CI -1 .66-3 .20 ,P=0 .53) ,number of positive re-moved lymph nodes(WMD=0 .08 ,95% CI -0 .23-0 .40 ,P=0 .61) ,postoperative drainage time(WMD= -1 .30 ,95% CI -6 .40 -3 .80 ,P=0 .62) ,postoperative hospital stay (WMD= -4 .02 ,95% CI -10 .19-2 .15 ,P=0 .20) ,but the difference between VEIL and OIL in term of surgical complications had statistical significance (OR=0 .08 ,95% CI 0 .03-0 .26 ,P<0 .01) .Conclusion VEIL has equivalent efficacy to OIL ,but has less surgical complications .
10.Observation of clinical curative effect in the patients with internal endometriosis by interventional therapy
Peng WANG ; Xin-yan NG ZHA ; Shuai WANG ; Shun-ji SUN ; Xiu-chun WANG ; Zhao-cheng JIAN ; Ye-quna SUN
Chinese Journal of Postgraduates of Medicine 2012;35(3):8-10
ObjectiveTo explore the curative effect in the patients with internal endometriosis by interventional therapy.MethodsUsing Seldinger technique,34 cases with internal endometriosis wereperformed bilateral uterine artery embolization.Observed postoperative menstrual quantity,dysmenrrhea degree,anemia and the change of the volume of uterine lesions.ResultsAll the patients were followed up for 1-3 years,menstrual quantity average decreasd of 59.1%P < 0.05 ),the symptoms of dysmenorrhea was significantly eased in 28 cases (82.4%,28/34).All the patients of anemia haemoglobin were back to normal,volume of uterus average reduced 43.8%P < 0.05 ),lesion was obviously smaller or disappear.Ultrasonography showed myometrium and blood flow signal of lesion was was obviously reduced.Conclusion Internalendometriosis by interventional therapy can get good results,symptoms improve significantly.