Objective:In order to establish ELISA method for testing tobacco mosaic virus (TMV) infecting Zingiber officinale Rosc.Methods: Tobacco mosaic virus(TMV) infecting Zingiber officinale Rosc was tested by RT-PCR.Zingiber officinale Rosc leaves which only contained TMV were choiced.TMV particle was purified by centrifugation method.TMV CP was purified through preparation electrophoresis including 12％ SDS-PAGE first and then 5％-20％ gradient SDS-PAGE.Polyacrylamide gel contained TMV CP was ground into suspension.Mice were immuned with the suspension and antiserum was obtained.Antiserum quality was tested by Western blot and ELISA test.IgG was purified through affinity chromatography method.IgG solution was concentrated and dialyzed to a suitable concentration.The IgG then mixed with glycerol.Results: IgG in antiserum only combined with TMV CP protein and it could combine with nature TMV particle CP protein.Its quality was up to standard.Conclusion: Establishment of ELISA method for testing TMV infecting Zingiber offcinale Rosc is successfull by using this IgG.

Objective:In order to look for a good method for preparation of hemorrhagin antiserum. Methods: Three kinds of hemorrhagins including AaH Ⅰ, AaH Ⅱ, and AaH Ⅳ were purified from Agkistrodon acutus venom according to predecessors's methods and crude AaH Ⅰ, AaH Ⅱ and AaH Ⅳ were obtained. Preparation electrophoresis was used to purify AaH Ⅰ,AaH Ⅱand AaH Ⅳ further. As for an hemorrhagin, six different dyeing methods were used to dye PAGE gel and the gel contained hemorrhagin was obtained respectively. The ground gel contained hemorrhagin was used to immune mice and its antiserum was obtained. Antiserums quality was tested through ELISA test and neutralization of the hemorrhagic activities of corresponding hemorrhagin. Results:Effective IgG concentration in different antiserum was different and effective IgG made through non toxic type protein fast stain reagent kit was higher than others. Conclusion:Non toxic type protein fast stain reagent kit is the best dyeing method among the six dyeing methods.

Objective To investigate the results of different measuring procedures of hemoglobin A1c (HbA1c) trueness verification scheme in China.Methods Cross sectional survey.The data were collected via the External Quality Assessment (EQA) software from laboratories participated in the First HbA1c trueness verification EQA.Then the collected data were divided into several groups based on laboratory instruments and the data from less than 5 group were excluded.The observed imprecision, bias and sigma (σ) were calculated and the bias％ and CV％ were drew in the sigma chart.The average bias％, CV％ and weighted average σ of each level were also calculated.Results Total 123 laboratories were divided into 9 groups and setting 6％ as the Allowable Total Error, the average bias％, CV％ and weighted average σ of 201411 (target value was 5.4％) were 3.70％, 4.55％ and 0.51 respectively σ, of 201412 (target value was 7.8％)were 2.42％ , 3.56％ and 1.24σ respectively.None of the group achieved the 2σ quality of 201411, and 1 group achieved the 2σ quality of 201412.Conclusions There are obvious biases among the results of many measuring systems and the target value assigned by reference measuring procedures of HbA1c, as well as the imprecision.The Sigma External Quality Assessment Chart is a visual tool, indicating that the quality of measuring systems necessitate improvement therefore to ensure the reliability of results and make better use of HbA1c in clinical application.

Objective To determine the normal reference values for thromboelastography (TEG) in Chinese healthy elderly people in Beijing and compare them with those provided by the manufacturer and with people under the age of 55 years.Methods The healthy elderly people aged over 65 years were enrolled from March 2014 to April 2014.4 ml of peripheral vein blood samples were taken from each person.All subjects underwent the laboratory tests of prothrombin time (PT),activated partial thromboplastin time (APTT),thrombin time (TT) and fibrinogen (FIB).TEG parameters including reaction time (R),coagulation time (K),angle (alpha) and maximum amplitude (MA) were determined by using a Heamoscope 5000 device.Results A total of 119 healthy elderly subjects were enrolled in this study.The mean values (95％ CI) of TEG parameters for them were R:6.29 min (4.4-8.2 min),K:1.63 min (0.8-2.5 min),α-Angle:66.09° (55.0-77.1°),MA:63.43mm (54.5-72.3 mm).There were significant differences in the values of R,K and α-Angle between the elderly and non-elderly groups (P＜0.001 or 0.05).There were significant differences in the values of K,α-Angle and MA between males and females (P＜0.05 or 0.001).According to the normal reference values provided by the manufacturer,16.1％ (19/119) of the healthy elderly volunteers had at least one abnormal parameter beyond its normal range.3％ (3/119) of the healthy elderly people were diagnosed as coagulation abnormalities,yielding a specificity of 84％.Compared with normal reference values provided by the manufacturer,the value of R was lower and the value of MA was higher in the healthy elderly people in Beijing.Conclusions The normal reference values for TEG should be established based on the local population characteristics.

To study the long-term toxicity of arginine esterase from Agkistrodon halys ussuriensis venom for the clinical application of arginine esterase in the future. Beagle dogs were used as experimental animals and were divided into control group, arginine esterase low dose group (0.06 u/kg), the middle dose group (0.18 u/kg) and high dose group (0.36 u/kg). Every group consisted of four dogs. The arginine esterase was given intravenously once a day for 180 days. Then three dogs in each group were sacrificed and the fourth one was fed without injecting arginine esterase for 15 days. The toxic reactions during treatment and recovery period were determined by evaluating and comparing the general criteria ( including locomotor activity, growth rate, appetite and death rate), clinical criteria (including blood test and urine test), pathological dissection and viscera coefficient of the treated animals and the control animals. There were no significant differences in general criteria. The clinical criteria of the treated animals were the same as those of the control animals except liver function. There were no significant differences in pathological dissection and viscera coefficient between the treated animals and the control animals except livers. The livers in high dose arginine esterase treated animals were swollen and vacuolated and there was significant difference in liver coefficient between them (P<0.05). The toxic symptom of liver disappeared after withdrawal of treatment. From these results, the non-toxic dose of arginine esterase for dogs was estimated to be 0.18 u/kg under the present study conditions and is about 15 times the clinical dosage for using the drug "Qingshuanmei" of which the main component is arginine esterase. The long-term toxicity test result indicates that the toxicity of pure arginine esterase is lower than that of "Qingshuanmei", suggesting that clinical use of the arginine esterase is safe.
Animals ; Anticoagulants ; toxicity ; Carboxylic Ester Hydrolases ; toxicity ; Crotalid Venoms ; enzymology ; toxicity ; Dogs ; Female ; Kidney ; drug effects ; pathology ; Liver ; drug effects ; pathology ; Male

Objective To discuss the clinical effect of Sky bone expander percutaneous ky-pbeplasty (Sky-PKP) in treatment of old osteoperotic vertebral compression fractures. Methods The study involved 27 patients (27 vertebrae) with old osteoporotic vertebral compression fractures treated by Sky-PKP from March 2005 to June 2007. Normotopia, lateral and dynamia radiographs, CT scanning and MRI were performed preoperatively to verify fluid collection in the vertebral body, vacuum phenomenon and open-close phenomenon. Visual analog scale (VAS), Oswestry disability index (ODI), anterior body height, middle line body height, posterior body height and kyphotic angle changes were measured on a lateral radiograph before and after treatment. Results All patients were followed up for mean 6.2 months, which showed no severe complications. VAS score was decresed from preoperative 7.8 to postop-erative 3.1 and ODI from 65% to 37%. However, The anterior vertebral height and middle line vertebral height were recovered for 4.6 nun and 5.7 mm respectively compared with preoperation. Correction of ky-photic angle was mean 5.6°postoperatively. There was no sitatistical changes in regard to posterior body height before and after operation. Conclusions Sky-PKP is a reasonable procedure for treatment of old osteoporotic vertebral compression fractures under strict control of indications, especially with vacuum phenomenon, open-close phenomenon and fluid collection. While high degree of difficulty in puncturation results in insignificant correction of kyphotic angle and body height.

Objective To investigate the current situation of precision of internal quality control (IQC) in total cholesterol,triglyceride,HDL-cholesterol,and LDL-cholesterol and provide improvement measurements.Methods Web-based External Quality Assessment (EQA) system was used to collect IQCdata of lipid tests from 581 EQA participant laboratories nationwide.The data include the coefficient of variation (CVs) of IQC data under control in April 201 1 and long-term cumulative data.Excel 2007 was applied for data processing after excluding the invalid data.Acceptable rates of CVs of two-lot internal quality controls in 4 lipid testing were calculated according to 6 criteria,that were 1/4TEa,1/3TEa,allowable imprecision of National Cholesterol Education Program (NCEP) and the specifications based on biological variation including the optimal,appropriate and minimal allowable imprecision.Results Four hundred and thirty-five,434,405 and 360 laboratories reported the data of level 1 IQC for total cholesterol (TC),triglyceride (TG),HDL-C,LDL-C respectively,while 214,214,192 and 171 reported the data of level 2 IQC respectively.Acceptable rates of TC,TG,HDL-C,LDL-C based on NECP criteria were 69.2％ (304/435),85.3％ (370/434),61.3％ (48/405) and 69.0％ (248/360) for level 1 respectively while 81.3％(174/214),91.6％ (196/214),75.5％ (145/192) and 81.3％ (139/171) for level 2 respectively.In the group which met the NECP criteria,the proportion of using matching detection system was much higher than the group which did not meet the criteria.Conclusions It is an effective way for clinical laboratories to improve test quality by monitoring the current and cumulative CVs of internal quality control and comparing them against proper evaluation criteria to evaluate if the analysis system can meet quality requirements.

External quality assessments play important roles in quality improvement in clinical laboratories, but most laboratories focused on the unsatisfied data only.With appropriate quality controls, laboratories can detect not only the error sourses of unsatisfied data but the potential error sourse of satisfied data as well.

Objective To study the applicability of a new analytical specification defined in WS/T 403-2012 in the external quality assessment schemes and the external comparision of internal quality control .Methods It was a quality management method study.Total error allowable criterions listed in WS/T 403-2012 and GB/T 20470-2006 were selected to assess the results of 23 analytes in the 1st challenge of 2013 routine chemistry external quality assessment.The acceptable rate of 23 analytes were calculated with the two specifications.Criterions of imprecision derived from the two standards were applied to assess the coefficient of variation with internal quality control data.Results With the specification based on WS/T 403-2012, the ratio of laboratories that all five samples were passed in the 1st challenge for 23 analytes ranged from 55.5%to 94.7%.The ratio of laboratories with 80%or more samples passed in 2013 EQA ranged from 73.9%to 98.5%.While ratios of two kinds described above evaluated based on GB /T 20470-2006 ranged from 63.0%to 99.2%, and from 90.0% to 99.7%, respectively.The acceptable rate of CV according to the two criterions ranged from 55.5% to 94.7% and 63.0% to 99.2%, respectively.Conclusions As evaluation criterions of external quality assessment allowable total error and internal quality control imprecision in routine chemistry , the specification in WS/T 403-2012 can be used to assess the analytical performance of clinical laboratory more objectively and comprehensively.It can help laboratories to identify the latent problems for further quality improvement.

Objective To verify and monitor the performance of accuracy, precision and comparability of 26 clinical biochemical analytes (29 methods) in the six centers involved in multi-centers reference intervals research, and to ensure the reliability of theirmeasurement results.Methods During the period of the systems evaluating, two levels of commercial quality control materials and fresh frozen human serum reference materials were applied to verify the performance of inter-laboratory precision and accuracy of analysis systems. During the period of samples testing, the commercial quality control materials were measured whenever samples were analysed, the fresh frozen serum reference materials were measured once a month.The coefficient of variations (CVs), bias and total errors were calculated to assess the precision, accuracy and comparability.Results Verification of precision and accuracy: ( 1 ) the ranges of CVs of 29 methods in the six laboratory laboratories were 0.4%-6.0%, the CVs of all 29 methods met the criterion . (2) The overall average bias of the analysis systems of 21 analytes (24 methods) ranged from -5.15%( ALT) to 4.46% ( Ur ) .Among 24 methods the overall average bias of TP, Glu-GOD, Ur, Cl, Ca exceeded the acceptable range.The quality assessment during the period of samples testing:(1) The overall average bias ranged from -1.95%(Ca) to 2.92%(Ur), median 1.26%, they all met the requirements of relevant standards.( 2 ) When commercial control materials were tested, the requirements of CVs were fulfilled for most methods in the six laboratories,and the CVs of TP, Alb, Cl, Ca exceeded the acceptable range.The overall average TE of all methods met the quality specification for the C-N controls material.For the C-P control material, only the overall average TE of TP (5.05%) exceeded thearceptable range while the other methods met the requirement in criterion.Conclusions The performance of precision and accuracy of the analysis systems used in the six laboratories passed the verification.During the period of sample testing, the performance of precision and accuracy of the most methods in the 6 laboratories met the requirements of quality specifications, and the overall performance was good.Because of the limitation of current technology the performance of some methods didn't fulfill the requirement of specifications, and need to be improved.