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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.
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ObjectiveTo establish the proper review rules for the microscopic screening of urine samples tested by automatic urinalysis work station which is composed of LabUMat urine dry chemical analyzer and Urised urine sedimental analyzer.Methods The paired comparison was used to analyze the results tested by microscopy and Urised.A total of 2015 random urine samples were enrolled to establish and validate review rules.All the samples were collected from the inpatients and ontpatients of General Hospital of the People's Liberation Army from May to November 2011 and tested by urinalysis work station.2015 urine samples were firstly tested by urinalysis work station,including both urine dry chemical analyzer and urine sediments analyzer.Then each urine sample was examined microscopically by two technicians-in-charge using double-blind method.The average results from the two technicians were used as review results.Compared with review results,we set up the review rules and evaluated the Irue positive rate,false positive rate,true negative rate,false negative rate (omission diagnostic rate) and review rate.According to different test methods by automatic urinalysis work station,four microscopic review protocols were defined:( 1 ) Protocol 1:based on chemistry results only,microscopy review was performed when any of WBC,RBC,PRO and NIT was positive; (2) Protocol 2:based on urine sedimental analysis only,microscopy review was performed when any of WBC,RBC and CAST count was over upper limit of the reference range ; (3) Protocol 3:if any of BLD vs.RBC,LEU vs.WBC was different between two systems,or quantitative results had two or more than two gradient differences,microscopy review was performed; (4) Protocol 4:if any of BLD vs.RBC,LEU vs.WBC was different between two systems,or CAST was over upper limit of the reference range,or alam appeared,microscopic review was performed.300 randomly selected urine samples were tested to validate the review rules.Omission diagnostic rate and review rate were used to evaluate the rules.Results According to our review rules,the positive samples rate was 41.14％ (829/2015) and the negative rate was 58.86％ ( 1186/2015 ) ; Positive samples were composed of RBC (50.30％),WBC (53.32％) and CAST (3.74％).The review rates of four protocols were 42.93％ (865/2015),39.70％ (810/2015),29.58％(596/2015),18.91％ (381/2015 ),respectively.The false negative rates (omission diagnostic rates) were 6.36％ (128/2015),4.42％ (89/2015),1.34％ (27/2015)and 1.04％ (21/2015)respectively.Protocol 4 was selected as an ideal plan.Additional 300 urine samples were tested using protocol 4 in order to confirm the review rule.The review rate,consistency rate,true positive rate,false positive rate,true negative rate,omission diagnostic rate were 19.67％ (59/300),91.67％ (275/300),35.67％ (107/300),7.67％(23/300),56.00％ (168/300),0.67％ (2/300),respectively.After image review revised,the review rate was 8.67％ (26/300).ConclusionThe review rules established by our research for Urinalysis Work Station can find the abnormal urine samples effectively and exactly and can reduce the workload significantly.(Chin J Lab Med,2012,35:810-814)
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Objective To establish the proper review rules for the microscopic screening of urine analyzed by UF-1000i automatic urinalysis work station (composed of UF-1000i urine flow cytometer and AX-4030 urine dry chemical analyzer).Methods A total of 2 839 random urine samples were collected at Chinese People′s Liberation Army General Hospital from September 2009 to February 2010, and were analyzed using UF-1000i urinalysis work station.The parameters obtained from UF-1000i and AX-4030 included RBC, WBC, CAST and ERY, LEU, PRO.After analysis by urinalysis work station, each urine sample was examined microscopically by two technologists using double-blind method.The average results got from the two technologists were regarded as the judging criterion.Based on the criterion, the review rules for the 2 839 urine samples tested by urinalysis work station were created and adjusted, and the true positive rate, false positive rate, true negative rate, false negative rate and review rate of these review rules were calculated.After that, 299 randomly selected urine samples were tested to validate these review rules.Omission diagnostic rate and review rate were used to assess the clinical practicability of the review rules.Results Thirty seven rules for microscopic review and twenty seven rules without further microscopic examination were set up based on six parameters using UriAccess 3.0 Software.The microscopic examination result was taken as the judging criterion, the consistency rate of these rules was 81.11%(2 311/2 839), the true positive rate was 40.51%(1 150/2 839), the false positive rate was 16.17%(459/2 839), the true negative rate was 41.00%(1 164/2 839), the false negative rate(omission diagnostic rate) was 2.43%(69/2 839) and the review rate was 18.28% (519/2 839).Additional 299 urine samples were assayed using UriAccess3.0 software to further verify these review rules.The consistency rate was 82.27%(246/299), the true positive rate was 36.12%(108/299), the false positive rate was 16.39%(49/299), the true negative rate was 46.15%(138/299), the false negative rate(omission diagnostic rate) was 1.34%(4/299), the review rate was 19.06%(57/299). The 4 false negative samples selected by these review rules did not come from the nephropathy department or the urology department.Microscopic results of RBC and WBC form these 4 samples ranged 3-8 cells/HP. Thus, these review rules could avoid the missed diagnosis of those patients with severe renal dysfunction.Conclusion The review rules established from this study for the UF-1000i urinalysis work station can effectively detect abnormal urine samples and improve the efficiency and the quality of urinalysis in routine clinical practice.
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Base on the situations of urine analysis at home and abroad and the wrong tendency of neglecting the urine formed elements in routine test, the authors describe the clinical value and standard test protocol according to the national guideline, references, personal clinical practice and research findings. The authors also make comments on the advantages and disadvantages of microscopic analysis with a variety of instruments, and provide suggestions on how to strengthen the quality management of urine analysis in our couutry.
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Objective To determine the biological reference interval of urinary conductivity in healthy people and to study the relationship between urine conductivity and other parameters as well as its clinical feasibility.Methods Conductivities of midstream urine specimen from healthy people (n=10119,5074 males and 5045 females,aged under 96 years) or patients with different diseases (n=3449) were determined.Among them the following parameters:conductivity,osmolality,specific gravity,creatinine,urea,uric acid,sodium,potassium and chloride of 250 random urine specimens were simultaneously determined.Results The urinary conductivities in healthy people exhibited normal distribution and significant differences were found in the subjects with different age and sex.The reference range of urinary conductivity was between 10.42?4.61 mS/ch and 24.10?6.81 mS/ch in healthy people and between 7.95 mS/ch?2.40 and 18.01?5.90 mS/ch in the patients with different diseases.Conductivity determined was positively correlated with osmolality (r:0.894),specific gravity (r:0.727),sodium (r:0.698),potassium (r:0.563),chloride (r:758),uric acid (r:0.521),urea (r:0.556) and creatinine (r:0.495).Conclusions Urine conductivity,which determination is simple and rapid,may reflect the conductive capacity of electrolytes in urine and positively correlated with osmolality,so it can be used as a new parameter for urinalysis to diagnose renal concentrative function in routine laboratory.
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Objective To evaluate the accuracy and sensitivity of three techniques in the diagnosis of renal diseases following multiple myeloma. Methods 41 serum samples from the kidney-damaged patients with multiple myeloma and 36 from the control group with general renal diseases were detected by quantitative analysis of immunoglobulins, serum protein electrophoresis and serum Immunofixation Electrophoresis. The accuracy and sensitivity of the three techniques were analysed by Two-way ANOVA and Multiple Comparisons of the check-out rate of monoclonal immunoglobulin. Results No monoclonal components were checked out by quantitative analysis of immunoglobulins. The checkout rate of IgG and IgM myelomas were 100% by serum protein electrophoresis, which had application limit on other types of myelomas. Whereas all secretarial myelomas could be diagnosed and typied by Immunofixation Electrophoresis, the sensitivity and accuracy was 100%, there was no false positive in the control group. Comparing with quantitative analysis of immunoglobulins and serum protein electrophoresis, serum Immunofixation electrophoresis had higher sensitivity and accuracy in diagnosis of renal diseases following multiple myeloma ( P
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0.05).RBC and haematocrit(HCT)were significantly decreased(P