Objective To study the influence of Yin-nourishing Qi-tonifying and blood-activating recipe(养阴益气活血方) on anticoagulation and fabrinolysis of cultured human umbilical vein endothelial cells(HUVEC).Methods Yin-nourishing Qi-tonifying blood-activating recipe was composed of three equal proportions of Radix Rehmanniae(生地黄),Radix Astragali(黄芪) and Rhizoma Chuanxiong(川芎);blood-activating recipe(活血方)contains Rhizoma Chuanxiong only.Serology pharmacology method was used to observe the effects of Yin-nourishing Qi-tonifying and blood-activating recipe on anticoagulation and fabrinolysis of cultured endotheliocytes of human umbilical vein.Results Compared with control group,both Yin-nourishing Qi-tonifying blood-activating and blood-activating recipes could obviously increase the content of 6-keto-prostacyclin 1?(6-keto-PGF1?) in HUVEC((412.5?42.7)ng/L and(231.7?30.1)ng/L vs.(137.6?13.5)ng/L),promote the activity of tissue plasminogen activator(t-PA)((0.920?0.072)kU/L and(0.679?0.062)kU/L vs.(0.516?0.052)kU/L),and inhibit the activity of tissue plasminogen activator inhibitor(PAI)((0.622?0.071)kAU/L and(0.851?0.085)kAU/L vs.(0.934?0.076)kAU/L),decrease the content of endothelin(ET)((35.7?4.9)ng/L and(46.8?5.1)ng/L vs.(58.6?6.2)ng/L),and increase the content of nitrogen monoxide(NO)((21.68?2.26)mmol/L and(15.15?1.73)mmol/L vs.(8.67?1.24)mmol/L) of cultured HUVEC(P

ObjectiveTo evaluate the performance of antinuclear antibody (ANA) detection in clinical laboratories.Methods There were 2 external quality assessments (EQA) scheme for nuclear antibody detection.The panel consisting of 5 samples was distributed.Each participant laborotory of the EQA program was required to report the ANA qualitative results,patterns,titers and anti-double strain DNA (dsDNA) antibody,anti-extractable nuclear antigen(ENA) antibody,the percent agreements of which were calculated respectively.ResultsThe number of laboratories performing ANA test with IIF increased from 77.6％ ( 149/192 )in 2006 to 82.2％ (342/416) in 2011,while the number of laboratories performing ANA test with ELISA was in the range of 14.5％ ( 53/365 ) and 16.0％ ( 52/326 ).The positive percent agreements of IIF was over 98％.The positive percent agreement of ELISA were all over 90％.IIF showed more satisfying positive percent agreements than ELISA every year.Over 90％ of the laboratories reported correct results for samples with granular ANA pattern except 0613 and 0624.Over 95％ of the laboratories reported correct results for samples with homogeneous ANA pattern.Two samples with centromere pattern were correctly detected by 88.5％ ( 161/182 ),79.0％ ( 147/186 ) of the laboratories in 2007,while the sample with centromere pattern was correctly detected by 98.4％ (299/304), which indicated an improvement in the detection of centromere pattern.In ANA positive results,the lowest percentage of the laboratories reporting the median result was 36％ (94/261),while the highest percentage was only 85.5％(224/262).The satisfied results of anti-ENA antibody were over 90％.And those of anti-dsDNA antibody was over 85％.ConclusionsIIF is the most common method for ANA screening in clinical laboratories.ELISA is also used in some laboratories.The two methods reported satisfying results in ANA test.The detection of anticentromere antibodies is improved.But the results of ANA titer reported are unsatisfactory. ANA detection in routine practice needs to be improved by standardization.

Objective Anti-RNase virus-like particles containing HCV RNA 5'-UTR were used as positive samples in national external quality assessment ( EQA ) to evaluate the competency of clinical Laboratories for the quantification of HCV RNA and analyze the possible problems of domestic kits. Methods The quality control samples with target values in EQA panels were distributed nationally twice by National Center of Clinical Laboratory (NCCL) to participating laboratories for the quantification of HCV RNA in 2008 and 2009. Each panel consisted of 5 samples. All participants were required to carry out the detection and to return results in expected time. Positive samples were virus-like particles which had been calibrated against the WHO HCV International Standard (NIBSC96/798)and the results of positive samples from participants should be in the range of target value of logarithm ± 0. 5. The 2nd panel in 2008 contained the common HCV genotypes and the 2nd panel in 2009 contained serial diluted samples of genotype 1b. The results of positive samples detected with 3 different lots reagent (21001,21078 and 21097) from the 2nd EQA in 2009 were statistically analyzed using the analysis of variance, then Dunnett'S T3 and Tamhane'S T2 were used if heterogeneity of variance was found. Results There was 390 participating laboratories in 2008 and 428/426 in 2009. The percentages of laboratories within the range of target value of logarithm ± 0. 5 for varied genotypes were different. The percentages of laboratories for 1b were more than 91%, for 2a were 93.7% and 74. 2% ,for6 were 83.3% and 80. 3%. The CVfor the low-level sample was higher than that for the high-level sample in the same year. The numbers of laboratories reporting false-negative samples in 2008and the 2nd in 2009 were 5, 1 and 10 respectively. Statistical differences were found among the results of four quality control serum samples using 3 different reagents( F = 288.23, 324. 79, 291.98 and 261.16,P ＜0. 01 ). Conclusion The competency for detecting low concentration samples and samples with genotype 2a or 6 needs to be improved.

Objective To investigate the potential role of human cytomegalovirus lower matrix phosphoprotein 65 (HCMV pp65) in murine systemic lupus erythematosus (SLE).Methods The prokaryotic vector pET-28b and eukaryotic vector pcDNA 3.0 were constructed to express the HCMV pp65 protein.All the C57BL/6 mice were inoculated with pp65 eukaryotic vector intramuscularly five times at 2-week intervals and then were bled via the retro-orbital vein.Subsequently,indirect ELISA was used to evaluate the concentration of anti-pp65 IgG,anti-dsDNA and ANA.At the same time,IL-1 b,IL-6,and TNF-α were determined by competitive ELISA.Results The early onset of autoantibodies and an overexpression of IL-6 were observed in immunized male C57BL/6 mice.Conclusion HCMV pp65 triggers the deregulation of humoral immunity in C57BL/6 mice,which indicates that the immune responses induced by HCMV pp65 may be involved in the development of SLE.

Objective To evaluate the performance of HCV RNA detection in the first EQA program in 2012 and analyze possible problems in clinical laboratories.Methods The panel consisting of 5 samples was distributed to 927 laboratories.Each panel contains one negative sample and 4 positive samples,which were virus-like particles calibrated by international standard.The pere ent agreements of all the laboratories for qualitative and quantitative results were calculated.Genomic means (GM) and standard deviations (s) of all laboratories and each reagent were calculated.The overall GM and the GM of each reagent were compared with expected results and correlation curves were calculated.Results The percent agreements of sample 1211,1212,1213,1214 for qualitative results wcrc 99.5％ (403/405),98.5％ (400/406),100.0％ (405/405),100.0％ (406/406),respectively.The percent agreement of the negative sample was 99％ (401/405).The percent agreements of sample 1211,1212 and 1213 for quantitive results were similar,which were 93.8％ (549/585),92.3 ％ (541/586) and 94.5％ (554/586).However,the agreement of sample 1214 was only 87.7％ (514/586)and the agreement of sample 1214 for reagent A was 67.2％ (92/137).The overall GM agreed with expected results,while GMs of reagent C,E and G deviated from expected results.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent C were 4.22,3.56,5.16 and 5.90,respectively.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent E were 4.52,3.78,5.55 and 6.29,respectively.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent G were 4.83,4.36,5.72 and 6.56,respectively.Conclusions The overall results of HCV RNA qualitative and quantitative detection are satisfactory.However,some problems still exist,such as deviation of GM of some reagents,the interassay variability,systematic deviation and accidental deviation,which show that the quality of reagents should be improved.

Objective To investigate the association of hepatitis B virus(HBV) S gene quasispecies with the outcome of HBV infection.Methods Serum samples were collected from three chronic HBV carriers, three chronic hepatitis B and three chronic severe hepatitis B patients.All subjects were male and with HBV genotype C.HBV S gene was amplified, and 20 clones of HBV fragment were randomly selected and sequenced from each sample.SPSS 15.0 software was adopted for analysis.Results Quasispecies complexity of HBV S gene in chronic HBV carriers and chronic hepatitis B tended lower than that of the severe chronic hepatitis B, but the difference was not of statistical significance (P＞0.05).In T cell epitope 45, 47, 85 amino acid sites of the HBV S gene, the constitution of quasispecies in the chronic hepatitis B was more complex than that of the HBV carriers (P=0.01), but compared with the severe chronic hepatitis, the difference was not significant (P=0.06).The computer model showed that both the dominant clones and the non dominant clones could effectively bind to the receptors of cytotoxic T lymphocytes.Conclusion Quasispecies in some T cell epitopes of HBV S gene may be related with the clinical outcome of hepatitis B.

Objective To evaluate the agreement of antinuclear antibody (ANA) titer reported in clinical laboratories and analyze possible problems in clinical laboratories.Methods Experiment survey.The panel consisting of 5 samples was distributed to 533 laboratories.Each panel contains one negative sample and 4 positive samples,which were from individuals of autoimmune disease.ANA titer system was divided into traditional titer system with two-fold dilution and titer system with 3.2 times dilution.Clinical laboratories were required to report the ANA titers and initial screening dilution according to the standard used in routine work.Results were expressed as median titers and range for acceptable performance.As to titer system with two-fold dilution,acceptable performance on proficiency testing was defined by a result equal to median titer ± two two-fold dilutions.While as to titer system with 3.2 times dilution,acceptable performance on proficiency testing was defined by a result equal to median titer ± 3.2 times dilutions.The laboratories percentages with acceptable performance were calculated to evaluate their agreements.Results 412 laboratories reported ANA titer results,of which 11.9％ (49/412)reported results with two-fold dilution titer system,88.1％ (363/412)reported results with 3.2 times dilution titer system.The median titers of sample 1211,1212,1213 and 1215 reported with two-fold titer system were 1 ∶ 640,1∶ 320,＞ 1 ∶ 1280 and1∶ 160,respectively.The agreement within the median ANA titer reported with two-fold dilution titer system ranged from 24.5％ (12/49) to 57.1％ (28/49) and the lowest percentage of the results within the acceptable limits was 87.8％ (43/49).The median titers of sample 1211,1212,1213 and 1215 reported with 3.2 times dilution titer system were 1 ∶ 1000,1∶ 1000,＞ 1 ∶ 3200 and 1 ∶ 320,respectively.The agreement within the median ANA titer reported with 3.2 times dilution titer system ranged from 63.3％ (31/49)to 83.7％ (41/49).The lowest percentage of the results within the acceptable limits was 98.0％ (48/49).Conclusions The results of ANA titer reported are unsatisfactory.Standardization for reagent,microscopy,procedure and result interpretation is necessary to improve the agreement of ANA titer report in different laboratories.

OBJECTIVE To explore the phenotype and genotype of Klebsiella and Escherichia coli producing extended-spectrum ?-lactamases(ESBLs).METHODS The producing ESBLs strains in 110 E.coli and 94 Klebsiella isolates were determined according to disk diffusion test,recommended by NCCLS and the drug-resistant genes such as TEM,SHV,CTX-M-1,CTX-M-2 and CTX-M-9 were detected by PCR and sequencing.RESULTS The producing ESBLs rates were 36.2%,42.7% and 39.7% in Klebsiella,E.coli,and in both two strains,respectively.The positive rate in detecting ESBLs by CTX and CTX/CLAV was higher(66.7%);the negative rates were 68.1% in E.coli,64.7% in Klebsiella when using CAZ alone and CAZ/CLAV.The rates of TEM,SHV,CTX-M-1,CTX-M-2 and CTX-M-9 were 54.3%,38.3%,21.0%,24.7% and 70.4%,respectively,in producing ESBLs isolations.The CTX-M genotype was predominant,(91.4%);the isolations(71.6%) contained more than two resistance genes.CONCLUSIONS More attention should be payed on ESBLs producing E.coli and K.pneumoniae strains.

Objective To explore the effects of general anesthesia combined thoracic paraverte-bral block on postoperative pain and fast track single-port video-assisted thoracoscopic surgery (VATS).Methods Thirty patients,including male 20 and female 10,received single-port VATS were randomly and equally divided into two groups:group C received general anesthesia only,and group T received ultrasound-guided thoracic paravertebral nerve block combined with general anesthe-sia.Both groups did not use the patient-controlled analgesia,if insufficient analgesia happened (rest-ing VAS scores>4),than used dezocine intravenously as additional analgesia (a single-dose 5-20 mg, no more than 120 mg per day).The Ramsay scores at 1,4,8,12 h after the surgery and the mechani-cal withdrawal threshold on the day before the surgery,at 4,8,12,24 h after the surgery were recor-ded.The first time of post-operation pain feedback,the consumption of dezocine in the first 24 h after surgery,the incidence rates of side effects,the first time off-bed and the hospital stays were also re-corded.Results Compared with group C,the Ramsay scores at 8,12 h postoperatively in group T significantly decreased (P <0.05),and the mechanical withdrawal threshold at 4,8 h postoperatively significantly increased (P <0.05).The first time of post-operation pain feedback in group T was sig-nificantly longer than group C (P <0.05).The consumption of dezocine in the first 24 h after surgery significantly decreased in group T (P <0.05).The first time off-bed and the hospital stays in group T were shorter than group C (P <0.05).Also,the incidence rates of nausea,vomiting in the first 24 h postoperatively were lower in group T (P < 0.05 ).Conclusion General anesthesia combined with single-injected thoracic paravertebral nerve block can effectively relieve the postoperative pain in pa-tients undergoing single-port VATS,reduce the consumption of opioids in the first 24 h postopera-tively,cutting down the occurring rates of adverse reactions,which was beneficial to early ambulate and shortened the hospital stays.

Objective To develop a candidate reference method for the measurement of creatinine in human serum based on isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS). Methods An isotopically labeled internal standard [<'2>H<,3>] creatinine was added to the serum sample and equilibrated with the endogenous creatinine. The samples were treated with anhydrous ethanol to remove proteins by precipitation. After being washed with chloroform for further clean-up, the samples were analyzed by LC/MS/MS. Serum creatinine was quantified by a bracketing calibration. Results The within-run, between-run and total coefficients of variation ranged from 0.52% to 0.61%, 0.11% to 0.59% and 0.61% to 0.83%, and the averages were 0.57%, 0.43% and 0.73%, respectively. The analytical recoveries ranged from 99.09% to 101.13% with an average of 100.3%.The results of analyzing the certified reference material SRM 909b (Level Ⅰ and Ⅱ) and SRM 967b showed biases of less than 0.4%. Conclusions An ID-LC/MS/MS method for measuring serum creatinine has been developed. The method is highly precise and accurate and may be used as a candidate reference method for serum creatinine measurements.