In the clinical laboratory medicine,the measurement uncertainty (MU)is a relatively new concept.Over the years, experts of clinical laboratory medicine from all over the world made a great number of further researches and promote the development of MU,which led clinical laboratories to pay more and more attention to the meanings and functions of MU at the same time.However,because of the habitual using of the total error (TE)in clinical laboratories and similarities between concepts of MU and TE which easily resulted in confusion,a lot of laboratories still cannot completely accept MU.By explai-ning concepts of TE and MU and analyzing the pros and cons of models of TE and MU as well as their functions,the obj ec-tive of this paper is to help clinical laboratories make further comprehensions of TE and MU and understand how to properly use them in practice.

In clinical laboratory medicine,measurement uncertainty (MU) is a fixed property of testing results in the measuring system.As an important part of ISO 15189,it is necessary for clinical laboratories to determine MU during the period of validation and verification for each measurement procedure and to review MU over time.Now,testing reports provided by clinical laboratories usually do not offer MU,but some clinical laboratories have already estimated MU in their routine work.Estimation andmonitoring of MU can help clinical laboratories offering more accurate results and provide objective tools for clinicians used in result intcrpretatinn.Generally,result interpretation can be achieved by the result comparison with three main comparators,including a previous result from the same patient,a population reference interval and a clinical decision point.The means of true value and the components contributing to the estimation of MU are both different when the com parison is conducted between testing results and different comparators,so the optimum estimation method of MU is accordingly different,which will subsequently affect the MU value and the determination of clinical decisions.Obviously,depending on the actual clinical uses,laboratories can choose appropriate comparators to the result interpretation and the determination of optimum estimation method of MU.For different clinical uses (diagnosis or monitoring) of the same mearurands,the adoption of different estimation methods should be used to acq uire reasonable MU.By interpreting the concept,characteristics,estimation,and uses of MU,as well as explaining how three main comparison methods of results exploit their own traceable chain to get MU,this paper intends to help clinical laboratories get further understanding of the importancc of MU and provide guidance for the MU estimation in routine work.

Objective Discussthe reference intervals and their sources of lymphocyte subsets by flow cytometry tests by all Chinese clinical laboratories at 2014 with a comparison of difference between different test systems.Methods Questionnaires about reference intervals were distributed to 212 clinical laboratories participating in October 2014 national external quality assessment scheme of lymphocyte subsets by flow cytometry tests of National Center for Clinical Laboratory.Relevant information about reference intervals of 5 relative counting items CD3 + (％),CD3 + CD4 + (％),CD3 + CD8 + (％),CD3-CD16 + CD56 + (％) and CD3-CD19 + (％) was collected by a internet-based external quality assessment software systems.Microsoft Excel 2010 and SPSS 19.0 were used to sort out raw data,eliminate unreasonable or incorrect data,and only analyze source distribution,arithmetic average,median,minimum value,maximum value and percentiles (P2.5 and P97.5) of upper and lower limits about healthy adults' reference intervals.Laboratories derived from the highest percentage of source distribution whose reagent and instrument were matching with each other were grouped according to the test systems used and the differences of reference intervals between different systems were compared with Mann-Whitney test.Results For five items collected,in the clinical laboratories who accepted statistical analysis,respectively 39.32％ (81/ 206),37.44％ (79/211),37.68％ (78/207),37.11％ (72/194),36.98％ (71/192) of them had verified the reference intervals they used.The top three sources of reference intervals were instructions of reagent and instrument manufactures (49.70％-52.54％),calculations by laboratories themselves (18.64％-20.00％),and calculations by other laboratories (14.12％-15.43％).The arithmetic averages and medians of upper and lower limits about every item were relatively close.But the minimum and maximum values of upper and lower limits of certain items had moderate differences.As Mann-Whitney test later showed,all items with the P values of upper and lower limits between different test systems,except for the lower limit of CD3 + CD4 + (％) (P value is 0.052) and the upper limit of CD3-CD19 + (％) (P value is 0.117),were lower than 0.05 which meant an existing statistical difference.Conclusions The sources of lymphocyte subsets by flow cytometry tests in all Chinese clinical laboratories were different and the instructions of reagent and instrument had the highest percentage,but the reference intervals among differenttest systems had statistically significance.

Objective To explore the clinical application of digital chair?side immediate all?ceramic restoration system,design and fabricate inlays and onlays to restore the large?defected molars,and to analyze and evaluate the efficacy of this clinical restoration. Methods Totally 84 inlays and 65 onlays were fabricated with IPS e.max CAD series ceramic blocks by using Cerec AC system to conduct immediate all?ceramic restorations of 149 teeth for 108 patients. The inlays and onlays were applied and cemented to the prepared teeth with dual?cured resin luting agent. The restored molars were re?examined by two dentists after six months. The analysis standard was based on the United States Public Health Service(USPHS criteria) and the results were statistically analyzed(α=0.05). Results There was no statistically significant difference in the case numbers of grade A and grade B for each index comparing the immediate condition to the condition after 6 months. Over 90 percent of ceramic inlays and onlays were quali?fied to be grade A. The restorations were well in the aspects of anatomic morphology,marginal integrity,color match,marginal discoloration,reten?tion,gingival health and secondary caries. Conclusion Computer aided design/computer aided manufacture(CAD/CAM)system combined with machinable glass ceramics can achieve a favorable result in restoration of large?defected teeth. Digital chair?side immediate all?ceramic restoration technique is an ideal solution for large?defected tooth restoration.

Objective To analyze the reference intervals of serum total bilirubin (TBIL) and serum direct bilirubin (DBIL) by all Chinese clinical laboratories and make a comparison with the upcoming part 4 of Industry Standard WS/T 404.Methods Relevant information about reference intervals of all clinical laboratories participating in TBIL and DBIL testing items of 2014 national external quality assessment scheme of clinical routine chemistry was collected by a web-based external quality assessment software,including source,grouping,verification,upper and lower limits of reference intervals and instruments,reagents,methods and calibrators used.Microsoft Excel 2010 and SPSS 19.0 were applied to statistical analysis.The comparison between reference intervals used and the upcoming Industry Standard was conducted by the simple mean t test.Laboratories derived from the highest percentage of source distribution whose reagent and instrument were matching with each other were grouped according to the test systems used and the differences of reference intervals between three mainly used test systems and the upcoming Industry Standard were also compared with the single sample mean t test.Results The number of laboratories participated in the investigation about source distribution of reference intervals for TBIL and DBIL was 749 and 709 respectively.For these two items,the highest sources were both instructions of reagent(TBIL 58.08％,DBIL 58.67 ％),next were both National Clinical Laboratory Operation Rules (3rd Edition) (TBIL 29.64 ％,DBIL 28.91 ％),the percentages of other sources were all less than 10％.Besides,there were respectively 379 (50.60％) and 354 (49.93％) laboratories verifying their reference intervals for TBIL and DBIL.The difference of the comparison between reference intervals used and the upcoming Industry Standard had statistical significance (P＜0.05).The using reference intervals were narrower than the upcoming Industry Standard for both items.In all laboratories with reference intervals from instructions of reagent,themating laboratories respectively accounted for 41.88％ and 41.48％％ for item TBIL and DBIL.As the single sample mean t test showed,the comparison of the reference intervals between three mainly used test systems in these mating laboratories and the upcoming Industry Standard had statistical significance (P＜0.05),except for the lower limit of DBIL with Beckman test system (P value was 0.068) and the upper limit of DBIL with HITACHI test system (P value was 0.087).Conclusion Currently,the using situation of reference intervals about TBIL and DBIL by all Chinese laboratories was not scientific and rational enough and has significant difference with the upcoming Industry Standard.Should publish the part 4 of Industry Standard WS/T 404 as soon as possible,which would help clinical laboratories establishing suitable reference intervals and promote the standardization of its usage.

Objective To evaluate the effects of age factors on the pharmacokinetics of ropivacaine during combined lumbar plexus-sciatic nerve block in patients.Methods Twenty patients of both sexes,aged ≥ 18 yr,weighing 50-75 kg,of American Society of Anesthesiology physical status Ⅰ or Ⅱ,scheduled for elective lower extremity surgery,were divided into 2 groups (n=10 each) according to age:young and middle-aged group (18-59 yr) and elderly group (≥ 60 yr).Combined lumbar plexus-sciatic never block was performed using an ultrasonic instrument and a nerve stimulator.Lumbar plexus block was performed with 0.5％ ropivacaine 30 ml.Sciatic never block was performed with 2％ lidocaine 10 ml plus 0.75％ ropivacaine 10 ml.Before administration and at 5,10,15,20,30,45,60,120,180 and 360 min after administration,blood samples were collected from the radial artery for determination of the blood concentration of ropivacaine by high-performance liquid chromatography.The area under the concentrationtime curve,maximum concentration (Cmax),time to C terminal elimination half-life and clearance were calculated.Results Compared with young and middle-aged group,the blood concentration of ropivacaine was significantly decreased at 5-45 min after administration,the Cmax was significantly decreased,terminal elimination half-life was significantly prolonged (P＜0.05),and no significant change was found in area under the concentration-time curve,the time to C and clearance in elderly group (P＞ 0.05).Conclusion Age factors can affect the pharmacokinetics of ropivacaine during combined lumbar plexus-sciatic nerve block,and both absorption and metabolism of ropivacaine are slower in elderly patients than in young and middle-aged patients.

ObjectiveTo compare liver pathology changes of patients with Budd-Chiari syndrome (BCS) and intrahepatic portal hypertension (IPH) after portosystemic shunt surgery. MethodsFrom January 2010 to December 2011,liverbiopsy was taken during shunt surgery (9 BCS patients,4 IPH patients),and 6-9 months after surgery on follow-up.Collagen type Ⅳ ( Col Ⅳ ),procollagen m (PC Ⅲ ),matrix metalloproteinase (MMP-1),tissue inhibitors of metalloproteinase(TIMP-1) were tested using SABC (immuonohistochemistry) method,and HE staining to observe the morphology of liver tissue.Free portal vein pressure before and after shunt was measured. ResultsIn BCS group,Col Ⅳ,PC 1Ⅲ and TIMP-1expression downregulated after surgery (127 ±15) vs.(137 ±16),t =4.896,P-0.013; (115.2 ± 10.6) vs.(127.3±9.5),t=4.877,P=0.003; (119.2±11.3) vs.(131.2±l9.6),t=2.841,P=0.023.MMP-1expression did not change ( P ＞ 0.05 ),while MMP-1/TIMP-1was not significantly correlated with liver fibrosis (0.95 ±0.16) vs.(0.98 ±0.15),t =-0.710,P =0.504.In IPH group,the expression of Col Ⅳ,PCⅢ,MMP-1,and MMP-1/TIMP-1did not change significantly after surgery (P ＞0.05).Compared with that in IPH group the expression of PC Ⅲ,Col Ⅳ and TIMP-1downregulated significantly in BCSgroup (127±15) vs.(150 ±12),U=3.000,P=0.038; (115.2 ±10.6) vs.(128.1±2.8),U=2.000,P=0.023; (119.2 ± 11.3) vs.(131.4 ±2.5),U=3.000,P =0.038.By HE staining in BCS group there was significant intrahepatic congestion which alleviated after surgery.While in PHT group liver pathology did not change significantly after surgery.FPP in BCS and IPH patients significantly decreased after shunt surgery (25 ±8) vs.(41±8) cmH20,t=17.816,P=0.000;(31±8) vs.(45 ±9) cmH20,t =5.745,P =0.010 ). Drop of FPP of BCS group plays a key role in reversal of liver fibrosis.ConclusionsIn BCS group liver pathology improved after shunt surgery probably by removing the intrahepatic obstruction,but in IPH group liver pathology remained unchanged after shunt.

OBJECTIVE To investigate the effect of obstructive sleep apnea hypopnea syndrome(OSAHS) on the quality of voice. METHODS Electroglottography (EGG) method was used for detecting the vocal cords vibration, waveforms of electroglottography and voice parameters such as Jitter, Shimmer, normalized noise energy (NNE), contact quotient(CQ), contact index (CI), contact quotient perturbation(CQP), contact index perturbation(CIP), and the in 82 OSAHS patients and 53 healthy subjects. The voice parameters were analyzed with Dr.Speech software, RESULTS While in normal pronunciation, the Jitter, Shimmer, NNE, CQP and CIP were (1.037±1.493)%, (5.627±1.872)%, (-16.109±1.719)dB, (2.638±0.633)% and (166.789±30.492)% in 22 mild OSAHS patients, (1.578±0.362)%, (8.478±1.635)%, (-11.517±1.614)dB, (6.043±0.936)% and (255.210±91.224)% in 24 moderate OSAHS patients, and (2.138±1.384)%, (14.888±4.089)%, (-7.677±2.151)dB, (11.749±7.164)% and (431.233±259.266)%in 36 severe OSAHS patients respectively. They were all higher than those in 53 normal subjects as (0.517±0.303)%, (3.299±2.196)%, (-21.933±5.477)dB, (1.322±0.942)% and (58.834±131.849)%. The regularity of vocal fold vibration in all patients with OSAHS was seriously out of whack. EGG waveform was irregular. CONCLUSION The OSAHS patients have structural changes in their upper airway, it will cause non-physiological breathing and lead dry of the vocal cord mucosa and weak elasticity. Their EGG parameters will get higher and vibration will be irregular seriously. We should pay attention to the voice qualities of the OSAHS patients and let them receive treatment as early as possible.

Objective To knockout the MATP gene of mouse melanoma cell line B16F10 using CRISPR/Cas9 system,and to lay foundation for the functional study of MATP gene.Methods Specific primers of MATP were designed according to the report in http://crispr.mit.edu/ website.The primers were linked to pCAS9/gRNA1 vector.Then the positive vector was transfected into mouse melanoma B16F10 cells,and monoclonal cell lines were obtained by the infinite dilution method.After the genomes of different monoclonal cell lines were extracted and sequenced,the cell lines with MATP gene cleavage were screened,and the expression of MATP in these cell lines was verified by Western-blot analysis.Results Three MATP gene knockout cell lines were successfully obtained.The western-blot results showed that the cell lines did not express MATP protein.Conclusions The knockout of MATP gene in B16F10 cell line can be successfully achieved using the pCAS9/gRNA1 vector.

Objective To observe CT features of primary yolk sac tumor (YST).Methods Clinical data and CT findings of 31 patients with primary YST proved by pathology were analyzed retrospectively.Plain CT was performed in 31 patients,while contrast enhanced CT scanning was performed in 23 patients.Results The lesions in 19 patients located in the gonads,including ovaries (n=11) and testes (n=8).Other lesions in 12 patients located out of gonads,including sacrococcygeal region (n =7),anterior mediastinum (n =3) and vagina (n=2).The tumors were oval shaped in 20 patients,while irregular shaped in other 11 patients.Well-defined boundary was found in 20 patients,whereas ill-defined boundary was found in 11 patients.Fat and calcification were found in 2 patients with teratomas.Moderate to marked enhancement of the solid part of tumors were observed in 23 patients,loofahs enhancement were observed in 17 patients,the blood vessels were found in 18 patients,while delayed enhancement of coated edge was found in 21 patients.The rupture of tumor capsule was found in 4 patients.Conclusion CT manifestations of YST have certain characteristics,which can provide imaging diagnostic evidences.