Objective The ADVIA Centaur XP automatic chemiluminescence immunoassay system detec-tion performance of the thyroid hormones and antibodies for validation and evaluation.Methods With refer-ence to the American association of clinical laboratory standardization guide and other related documents,the ADVIA Centaur XP automatic chemiluminescence immunoassay system test 7 items thyroid hormones and an-tibodies(T3,T4,FT3,FT4,TSH,anti-TG,anti-TPO)of precision,accuracy,linear range and carry pollution rate for validation.Results Within the seven thyroid hormones and antibodies batch testing precision of CV and CV between batch precision between 1.51% -6.17% and 1.86% -6.17%.Is the average accuracy of bias of the largest testing project FT3(8.47%),but in the acceptable range,good correlation,correlation coefficient R of 0.994 or higher,Average bias <1/2 CLIA′88 TEa(12.5%).T3,T4,FT3,FT4,TSH,anti-TG and anti-T PO in 1.24-5.59 nmol/L respectively,and 60.07 -195.10 nmol/L,3.40 -22.87 pmol/L,14.59 -40.54 pmol/L,1.63-74.13 μIU/mL,60.10 -381.30 μIU/mL and 180.10 -531.50 μIU/mL range is linear,Carry pollution rate is between 0.04% -0.97%.Conclusion ADVIA Centaur XP automatic chemiluminescence im-munoassay system detecting thyroid hormones and antibodies results consistent with the data provided by the manufacturer,the test results are accurate and reliable,and can be used for the detection of clinical samples.

Objective: To evaluate the ability of matrix-assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF MS) in the homology analysis of Carbapenems-resistant klebsiella pneumonia. Methods: Twenty-one non-duplicated strains of Carbapenems-resistant klebsiella pneumoniae were isolated from Shandong Provincial Hospital affiliated to Shandong University during April 2011 and October 2013 in this study. Twenty isolates were from neonatal unit and one from cardiac surgery. The homology analysis of Carbapenems-resistant klebsiella pneumoniae was performed with pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and MALDI-TOF MS respectively. Results: The result of PFGE was consistent with MLST. The twenty-one CRKPN strains were divided into three groups by MALDI-TOF MS according to their relationship, 18 of them belonged to II group, and the homology was higher than 75%. From the analysis of protein mass spectra of 18 strains, the protein peaks were roughly the same. Thus, it was concluded that their relationship was close, and the results were basically consistent with the results of PFGE and MLST. The H13 strain with low homology (<60%) was different from the above strains, especially in the molecular weight 4365, 5381 and 6289.The PFGE analysis showed that the homology between H13 and other strains was 61%, and the MLST classification result was ST54. Conclusions MALDI-TOF MS can be used to identify CRKPN accurately and analyze its homology analysis more conveniently than other methods in clinical laboratory. MALDI-TOF MS has the potential to be used as an easy and rapid epidemiology typing tool for nosocomial infection investigation caused by drug-resistant bacteria.(Chin J Lab Med, 2018, 41: 589-595)

Paget's disease of bone (PDB) is a common metabolic bone disease that is characterized by aberrant focal bone remodeling, which is caused by excessive osteoclastic bone resorption followed by disorganized osteoblastic bone formation. Genetic factors are a critical determinant of PDB pathogenesis, and several susceptibility genes and loci have been reported, including SQSTM1, TNFSF11A, TNFRSF11B, VCP, OPTN, CSF1 and DCSTAMP. Herein, we report a case of Chinese familial PDB without mutations in known genes and identify a novel c.163G>C (p.Val55Leu) mutation in FKBP5 (encodes FK506-binding protein 51, FKBP51) associated with PDB using whole-exome sequencing. Mutant FKBP51 enhanced the Akt phosphorylation and kinase activity in cells. A study of osteoclast function using FKBP51V55L KI transgenic mice proved that osteoclast precursors from FKBP51V55L mice were hyperresponsive to RANKL, and osteoclasts derived from FKBP51V55L mice displayed more intensive bone resorbing activity than did FKBP51WT controls. The osteoclast-specific molecules tartrate-resistant acid phosphatase, osteoclast-associated receptor and transcription factor NFATC1 were increased in bone marrow-derived monocyte/macrophage cells (BMMs) from FKBP51V55L mice during osteoclast differentiation. However, c-fos expression showed no significant difference in the wild-type and mutant groups. Akt phosphorylation in FKBP51V55L BMMs was elevated in response to RANKL. In contrast, IκB degradation, ERK phosphorylation and LC3II expression showed no difference in wild-type and mutant BMMs. Micro-CT analysis revealed an intensive trabecular bone resorption pattern in FKBP51V55L mice, and suspicious osteolytic bone lesions were noted in three-dimensional reconstruction of distal femurs from mutant mice. These results demonstrate that the mutant FKBP51V55L promotes osteoclastogenesis and function, which could subsequently participate in PDB development.
Acid Phosphatase ; Animals ; Asian Continental Ancestry Group ; Bone Diseases, Metabolic ; Bone Remodeling ; Bone Resorption ; Femur ; Humans ; Mice ; Mice, Transgenic ; Osteitis Deformans* ; Osteoblasts ; Osteoclasts ; Osteogenesis ; Phosphorylation ; Phosphotransferases ; Tacrolimus Binding Proteins ; Transcription Factors

Objective To analyze the difference of laboratory test results between early-onset and late-onset severe preeclampsia and to investigate their clinical application values.Methods Totally 108 blood samples were collected from patients with severe preeclampsia who were diagnosed according to the Diagnostic Standard of Obstetrics and Gynecology(7th Edition) published by People′s Medical Publishing House,in Shandong Provincial Hospital affiliated to Shandong University from March to November 2016,which consisted of 64 early-onset severe preeclampsia before 34 weeks gestation(early onset group) and 44 late-onset severe preeclampsia after 34 weeks gestation(late onset group).In addition,42 women with normal pregnancies as the control group were selected.General clinical data were collected,and the blood sample was analyzed through detecting Hb,PLT,fibrinogen (FIB),D-dimer,AST,ALT,urea,creatinine (Cr),uric acid,CRP,urine protein.The tested results were analyzed and compared.Flow cytometry was used to analyze the proportion of T helper 1 cells(Th1) and T helper 2 cells(Th2),and the ratio of Th1/Th2 was also calculated.All data and F test were performed by use of statistical software SPSS19.0.Results The pre-pregnancy body mass index(29.55±4.49,30.66±5.13,26.62±3.17,F=9.829,P<0.05),diastolic blood pressure[(105.17±14.46)mmHg(1 mmHg=0.133 kPa),(99.80±12.56)mmHg,(74.36±8.42)mmHg,F=82.088,P<0.05],Hb[(123.22±14.38)g/L,(117.03±16.48)g/L,(112.62±11.24)g/L,F=7.133,P<0.05],urea[(6.56±2.36)mmol/L,(4.51±1.35)mmol/L,(3.04±0.87)mmol/L,F=51.733,P<0.05],Cr[(68.47±18.05)μmol/L,(61.37±14.37)μmol/L,(48.54±8.73)μmol/L,F=23.737,P<0.05],CRP[(7.68±8.76)mg/L,(5.88±6.03)mg/L,(3.56±2.41)mg/L,F=4.735,P<0.05],urine protein[(3.66±0.76)g/L,(2.20±1.05)g/L,(0.19±0.40)g/L,F=249.714,P<0.05]had a statistically significant difference among the early-onset,late-onset and control groups.The flow cytometry results demonstrated that the proportion of Th1 in early-onset group(19.83±3.04)was higher than that in both late-onset (14.49±2.79)and control groups(11.78±1.17),on the contrary,the result of Th2 was much lower(early-onset:1.02±0.12,late-onset: 1.11±0.12,control: 1.56±0.11),there was statistical significance among these three groups(Th1: F=135.110,P<0.05;Th2: F=293.687,P<0.05).Conclusions It′s necessary to real-time monitor the laboratory indicators,such as liver and kidney function,especially the immunologic function indicators for evaluating the disease of early-onset and late-onset severe preeclampsia and personal treatment,and for ensuring the health of mother and fetus and improving the prognostic of mother and fetus.

Objective To investigate the infection status of Helicobacterpylori (HP) in the physical examination population and the contents of pepsinogen Ⅰ (PGⅠ),pepsinogen Ⅱ (PGⅡ) and gastrin-17 (G-17) in serumand the association with HP infection.Methods In this study,781 cases of physical examination were included.Serum HP antibody typing,PGl,PGⅡ and G-17 were measured by Helicobacter pylori antibody kit and enzyme-linked immunosorbent assay.The levels of PGⅠ,PGⅡ,PGⅠ/PGⅡ and G-17 were analyzed by SPSS22.0 analysis system,and the difference of HP infection rate between HP infection group and non-infected patients and different types of infection was analyzed.Analysis of the infection rate and the difference of each age group.Results There was no significant difference between the two groups (x2 =0.284,P=0.594),and there was significant difference between different age groups (x2 =8.523,P=0.014).The levels of PGⅠ,PGⅡ,PGR and G-17 in serum were statistically significant (Z=8.616～ 14.125,P=0.000) compared with those of HP positive and negative.There were significant differences in serum PGⅡ and PGR levels between HP Ⅰ type and HP Ⅱ type infection (Z=3.444,3.385,P=0.001).The levels of PGⅠ and PGⅡ in serum were significantly higher than those in other groups (Z=5.012,4.478,P=0.000).PGⅠ and PGⅡ were higher in women than in females (Z=0.444～0.941,P＞0.05),with the increase of age,PGⅠ,PGⅡ and G-17 has an upward trend,while PGR showed a downward trend.Conclusion The level of serum PG was closely related to gender,age and HP infection,and was closely related to HP infection classification.The combination of HP antibody typing and serum PG and G-17 as a routine test of gastric function is of great significance in the screening and evaluation of early gastric diseases.

Objective To verify and valuate the performance of small dense low-density lipoprotein cholesterol (sdLDL-C) detection by the direct clearance method and evaluate its preliminary clinical application in acute coronary syndrome (ACS).Methods Case control study:The performance (accuracy,precision,linearity) of sdLDL-C was assessed by direct clearance method.In 143 cases of ACS patients selected from Cardiology Department and Emergency Department of Shangdong Provincial Hospital from April to October in 2016,with 100 cases male,female 43 cases,including acute myocardial infarction (AMI)group of 59 cases,unstable angina pectoris (UAP) group of 84 cases;83 cases of healthy volunteers as a control group selected from health physical examination center of Shandong Provincial Hospital,with 59cases male,female 24 cases.Levels of sdLDL-C,total cholesterol (TCH),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),apolipoprotein A (ApoA I),apolipoprotein B (ApoB),lipoprotein (a) (Lpa) and high sensitive C-reactive protein (Hs-CRP) were detected separately by automatic biochemical analyzer.Non high density lipoprotein cholesterol (non-HDL-C) equals TCH minus HDL-C.x2 test,t test,one-way ANOVA,Pearson correlation and multiple linear regression analysis were used as statistical methods.Results The within-lot or between-lot variation was 2.85％ and 3.36％.Methodological comparison:regression equation Y =0.984X + 0.018,r2 =0.966,t =-0.191,P =0.850.There was a good linear correlation (Y =1.026X + 0.007,r2 =0.999) between theoretical values and actual detection results in range of 0.15-2.65 mmol/L.SdLDL-C concentrations were positive correlated with TCH,non-HDL,LDL-C,TG,ApoB (r =0.758,0.848,0.839,0.514,0.885,respectively,P ＜0.01),and negative correlated with HDL-C (r =-0.224,P =0.001),but no correlation with APOA I,Lpa and Hs-CRP(r =-0.021,0.050,0.003,respectively,P ＞ 0.05).Multiple linear regression analysis showed that the factors influencing sdLDL-C level were HDL-C,ApoB,LDL-C and TG.The levels of sdLDL-C,TG in the ACS group were significantly higher than those in the control group (t =3.415,4.660,respectively,P ＜ 0.01),but no difference between the two groups in the levels of TCH,non-HDL-C and LDL-C (t=-1.831,-0.452,-1.398,respectively,P ＞0.05).Comparing AMI group with control group,sdLDL-C,TG and Hs-CRP were significantly higher than the control group (P =0.000,0.000,0.000,respectively),but TCH,LDL-C and non-HDL were similar between the two groups (P =0.800,0.320,0.120,respectively);Comparing UAP group with control group,TG and Hs-CRP were higher than control group (P =0.001,0.047,respectively),TCH and LDL-C were significantly lower than the control group (P =0.003,0.008,respectively),but sdLDL-C had no difference (P =0.305);Comparing AMI group with UAP group,sdLDL-C,TCH,LDL-C and Hs-CRP were significantly higher than UAP group (P =0.000,0.003,0.001,0.000,respectively),and TG were no statistical significance (P =0.473).Conclusions Direct clearance method can meet the requirement of sdLDL-C detection.sdLDL-C level can assess the metabolism of blood lipids and be used as an independent risk factor and predictive index of ACS,superior to LDL-C.

Objective To investigate the clinical significance of Epstein-Barr virus EBV DNA in children with Epstein-Barr virus infection realated diseases.Methods A retrospective cohort study was performed.Totally 222 blood samples were collected from children who were diagnosed as EBV infection in Shandong Provincial Hospital from June 2012 to August 2013.Fluorescent quantitative PCR( FQ-PCR) was used to analyze the EBV DNA in peripheral blood lymphocytes.ELISA was used to analyze the four EBV serology antibodies in the serum.Two groups of tested results were compared.Heart, hepatic impairment and renal function were analyzed through detecting AST, ALT, BUN, CREA, CK, CKMB.The results were grouped by EBV DNA copy number, and then non-parametric test together with correlation analysis was performed using SPSS21.0 analytics software.Results The positive rate of EBV-CA IgM and EBV DNA was 51.35%(114/222) and 72.97% (162/222) respectively, χ2 =24.01, P<0.001.The EBV DNA copy number was significant difference (χ2 =11.79,P<0.05) in children at different stages of infection ( no previous infection:6.30 ×103 -1.20 ×104 copies/ml;early infection:5.56 ×103 -3.92 ×106 copies/ml;acute infection:6.58 ×103 -1.73 ×106 copies /ml;chronic infection or recurrent infection:8.92 ×103 -2.34 ×104 copies/ml;late infection or recovery:5.20 ×103 -1.12 ×107 copies/ml;past infection:5.46 × 103 -1.33 ×104 copies/ml).Each biochemical targets were divided into five groups by EBV DNA copy number (Ⅰ>1 ×106 copies/ml,Ⅱ1 ×105 -1 ×106 copies/ml, Ⅲ1 ×104 -1 ×105 copies/ml, Ⅳ5 × 103 -1 ×104 copies/ml, Ⅴ<5 ×103 copies/ml), and ALT(χ2 =10.14,P<0.05), BUN(χ2 =18.17, P<0.05), CK(χ2 =13.09,P<0.05), CKMB(χ2 =17.93,P<0.01) had a statistically significant difference between each group.Well, the log value of EBV DNA copy number had a positive correlation relationship with AST(r=0.357,P=0.001), ALT(r=0.376,P=0.001), BUN(r=0.329,P=0.000), CK(r=0.235,P=0.035).Conclusions Detection of EBV DNA can be used for the early diagnosis and assessment of process of the EBV infection related disease in children.The detection of liver, kidney function and myocardial enzymes can be used for evaluating the severity of EBV infection.

Objective To investigate the change and clinical value of serum procalcitonin (PCT ) in the patients with severe dia‐betic foot .Methods The serum PCT level was detected before treatment and at 1 ,2 weeks of treatment in 86 patients with severe diabetic foot .Results With the infection severity aggravating before treatment ,the elevation of PCT was more significant .The PCT level after 1 week treatment showed a decreasing trend ,but the difference was not statistically significant (P>0 .05);in the patients with Wagner grade 3-5 ,the PCT level after 2 week treatment showed a obvious decreasing trend ,moreover the difference was sta‐tistically significant(P<0 .05) .Conclusion PCT could serve as one of sensitive indexes to judge the outcome in the patients with diabetic foot .

Objective To investigate the antimicrobial resistant and transmission mechanisms of carbapenem-resistant K.pneumonia (CR-KP) infection of newborns.Methods A retrospective study was conducted on totally 37 non-repetitive CR-KP which were isolated from patients hospitalized between April 2011 and October 2013.Resistance genes were identified by PCR and sequencing.Plasmid was analyzed by pulsed-field gel electrophoresis (PFGE).Conjugation experiments were performed to determine the transferability of beta-lactamase.Multilocus sequence typing (MLST) was used to determine the genotypes and homology of these isolates.Out-membrane proteins were examined by PCR and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).Results Thirty-seven CR-KP isolates were tested.The resistant rates of imipenem, meropenem, ertapenem were 89.2％ (33/37), 83.8％ (31/37) ,97.3％ (36/ 37), respectively.All the 37 CR-KP exhibited 100％ (37/37) sensitivity to tigecycline, colistin, levofloxacin and amikacin, while resistance to most of the other antibiotics.By PCR, 67.6％ (25/37) isolates were blaNDM-1 positive, 35.1％ (13/37) isolates were blaIMP-4 positive and 2.7％ (1/37) isolate were blaIMP-8 positive, including two isolates carrying both blaNDM-1 and blaIMP-4.PFGE results showed that the isolates carried 2-4 plasmids and both blaNDM-1 and blaIMP-4 were transferable by plasmids.MLST assigned them to sequence type (ST) 20, ST17, ST54, ST705, ST290,which showed that there were infectious outbreaks caused by NDM-1-producing and IMP-4-producing respectively among newborns.SDS-PAGE result indicated that there was no absence of outer membrane proteins OmpK35 and OmpK36.Conclusions The main resistant mechanisms of CR-KP causing infection in newborns were those the isolates carried carbapenemase of blaNDM-1 or blaIMP-4 and the K.pneumonia with two kinds of carbapemenase were detected.

Objective To evaluate the usefulness of anti -HCV signal-to-cutoff (S/CO)ratio, AST, ALT and the combined examination of anti-HCV S/CO, AST, ALT for predicting HCV RNA results by a model of logistic regression and receiver -operating characteristic (ROC) curve.Methods Five hundred and eighty -eight anti-HCV positive samples were tested by ELISA , followed by RT-PCR to detect HCV-RNA and enzyme rate method to detect AST, ALT.Patients were divided into viremia and non -viremia groups according to HCV-RNA results.Logistic regression and ROC curve analysis was performed to evaluate the diagnostic accuracy of each index for a diagnosis of viremia.Results The serum anti-HCV S/CO ratio, AST, ALT of HCV-RNA positive group were higher than HCV-RNA negative group, showing significant statistical difference ( P <0.05).Anti-HCV S/CO, AST and ALT has a closely posi-tive correlation with HCV-RNA ( P <0.05), anti-HCV S /CO >AST >ALT.The Area Under Curve(AUC) of the combined examina-tion of anti-HCV S/CO ratio, AST and ALT was 0.949(95% confidence interval,0.932 to 0.966), higher than the AUC of anti-HCV S/CO ratio, AST and ALT single index examinations , which was 0.894(95 % confidence interval, 0.862 to 0.926), 0.823(95%confidence interval, 0.789 to 0.856) and 0.788(95% confidence interval, 0.750 to0.826 ) respectively.C onclusions The diag-nostic relevance of the three biochemical markers for predicting the presence of viremia was anti -HCV S/CO ratio >AST >ALT.The combined examination of anti-HCV S /CO ratio, AST and ALT in predicting hepatitis C viremia is superior to any single index examina -tion and it can increase the detecting ability of HCV -RNA greatly.