Application of improved comparability verification method in performance verification of instruments and equipment / 中国输血杂志

【Objective】 To explore the viability of improved comparability verification method for the performance verification of automatic biochemical analyzer in blood centers. 【Methods】 According to the relevant requirements of User Verification of Performance for Precision and Trueness; Approved Guideline-Second Edition (EP15-A2) and Guideline for Comparability Verification of Quantitative Results with One Health Care System(WS/T 407-2012), the performance of the newly purchased Beckman Coulter AU480 automatic biochemical analyzer was confirmed by means of comparability verification, and compared with the current Olympus AU400 automatic biochemical analyzer. 【Results】 The detection results given by Beckman AU480 biochemical analyzer on 5 samples for ALT detection performance test were significantly correlated with the target value (R2=0.998), and the trueness verification results met the requirements.The CV of medium and high ALT levels was 2.36% and 1.20%, respectively, within the acceptable standard range(<5%), and the precision verification results meet the requirement.The comparison deviation of two devices for medium and high ALT levels was 0.47% and 1.62%, respectively, within the acceptable standard range (<5%). The detection results of ALT from 40 blood donors given by two devices were significantly correlated (R2=0.956), and the precision verification results met the requirements.In the validation of linear evaluation, there was a good linear relationship between the expected and measured results (r=0.997)among samples with 6 levels of concentrations. 【Conclusion】 The comparability verification method is suitable for the performance verification of automatic biochemical analyzer in blood centers.

Activation of low-molecular-mass nitrile hydratase from Rhodococcus rhodochrous J1 by heterologous activators / 生物工程学报

As self-subunit swapping chaperones or metallochaperones, the activators assist nitrile hydratases to take up metal ions and they are essential for active expression of nitrile hydratases. Compared with nitrile hydratases, the activators have a low sequence identity. Study of the activation characteristics and the relationships between structures and functions of the activators is of great significance for understanding the maturation mechanism of nitrile hydratase. We co-expressed low-molecular-mass nitrile hydratase (L-NHase) from Rhodococcus rhodochrous J1 with four heterologous activators respectively and determined their activation abilities. Then we made sequence analysis and structure modelling, and studied the functions of the important domains of the activators. Results showed that all four heterologous activators could activate L-NHase, however, the specific activities of L-NHases were different after activation. L-NHase showed the highest specific activity after being activated by activator A, which was 97.79% of that of the original enzyme, but the specific activity of L-NHase after being activated by activator G was only 23.94% of that of the original enzyme. Activator E and activator G had conserved domains (TIGR03889), and deletion of their partial sequences resulted in a substantial loss of activation abilities for both activators. Replacing the N-terminal sequence of activator G with the N-terminal sequence of activator E, and adding the C-terminal sequence of activator E to the C-terminus of activator G could increase the specific activity of L-NHase by 178.40%. The activation by nitrile hydratase activators was universal and specific, and the conserved domains of activators were critical for activation, while the N-terminal domain and C-terminal domain also had important effects on activation.

Establishment and application of a new immune-comb assay for detection of serum antibody against simian immunodeficiency virus / 中国实验动物学报

Objective SIV30 protein of simian immunodeficiency virus(SIV)was prepared by genetic engineering technique as an antigen diagnostic reagent, to establish an immune comb method for the specific detection of anti SIV IgG in monkey serum. Methods Recombinant expression plasmid of SIV SIV30 gene was constructed by prokaryotic expression vector pGEX-4T-1, and expressed in the competent BL21 cells. The recombinant protein was purified as a diagnostic antigen, and a standardized procedure for the detection of immune comb was established and applied for clinical detection. Results The optimum coating amount of antigen was 0.02 mg/mL. The prepared IC was able to specifically detect the positive serum of SIV. There was no cross reaction between the sera of other viruses. It showed a high specificity of the detection method. Sensitivity analysis showed that the SIV30 protein was able to detect 1:400 times diluted SIV positive sera. The result of stability and repeatability test(the same sample was repeated 3 times) showed that the coefficient of variation(CV)was less than 10%. The serum samples of 10 suspicious monkeys were detected by this method, showing a consistent rate of comb method and ELISA test result of 100%, Kappa =1.000. Conclusions SIV30 protein is expressed in prokaryotic cells. The immune comb is prepared,and is successfullyl applied in clinical examination. It shows that the method has a high sensitivity, strong specificity, good reproducibility and practicability.

Biocatalytic access to β-alanine by a two-enzyme cascade synthesis / 生物工程学报

Enzymatic synthesis is an important way to produce β-alanine, but the biological method is expensive generally because of the high price of substrate. In this paper, a two-step enzymatic cascade system was developed, combining L-aspartase from Escherichia coli DH5α and L-aspartate α-decarboxylase from Corynebacterium glutamicum. This system catalyzes Fumarate and ammonia to β-alanine. The optimal ratio of AspA and PanD was 1:80 (W/W), and the concentration of AspA was 10 μg/mL, at 37 ℃ and pH 7.0. When the concentration of Fumarate was 100 mmol/L, the reaction reached its equilibrium after 8 h, and the yield of β-alanine was 90 mmol/L (7 g/L). The yield of β-alanine can reach 126 mmol/L (9.8 g/L) when the concentration of Fumarate increased to 200 mmol/L. Extending reaction time, the conversion rate did not change obviously. Using this two-step enzymatic cascade system, β-alanine from cheaper substrate Fumarate can be obtained.

Crosslinking sodium hyaluronate gel with different ratio of molecular weight for subcutaneous injection: animal experimental study and clinical trials subcutaneous injection / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the biocompatibility and degradation rate of crosslinking sodium hyaluronate gel with different ratio of molecular weight, so as to choose the effective, safe and totally degraded hyaluronate gel for aesthetic injection.</p><p><b>METHODS</b>(1) Compound colloid was formed by cross-linking the divinyl sulphone and sodium hyaluronate with different molecular weight (4 x 10(5), 8 x 10(5), 10 x 10(5), 12 x 10(5)). (2) Healthy level KM mice was randomly divided into two groups to receive hyaluronic acid gel or liquid injection. Each group was subdivided into three subgroup to receive hyaluronic acid with different molecular weight. The biocompatibility and degradation rate, of hyaluronate were observed at 7, 90, 180 days after injection. At the same time, different molecular weight of sodium hyaluronate gel is sealed or exposed respectively under the low temperature preservation to observe its natural degradation rate. (3) The most stable colloid was selected as aesthetic injector for volunteers to observe the aesthetic effect.</p><p><b>RESULTS</b>The sodium hyaluronate gel with molecular of 4 x 10(5) was completely degraded 90 days later. The sodium hyaluronate gel with molecular of 8 x 10(5) was completely degraded 180 days later. The sodium hyaluronate gel with molecular of 10 x 10(5) was degraded to 90.0% after 180 days. The sodium hyaluronate liquid can be degraded completely within 7 days. The colloid could be kept for at least 12 months when sealed under low temperature, but was totally degraded when exposed for I d. Sodium hyaluronate gel with molecular 10 x 10(5) was confirmed to be kept for at least 6 months in animal experiment and clinical trials.</p><p><b>CONCLUSIONS</b>Under the same condition of material ratio, the higher the molecular weight is, the lower the degradation rate is. But the liquidity of gel is not good for injection when molecular weight is too large. It suggests that Sodium hyaluronate gel with molecular 10 x 10(5) maybe the best choice in cosmetic injections.</p>

Therapeutic effect of comprehensive rehabilitation nursing on patients with cerebral infarction compli-cated type 2 diabetes mellitus / 心血管康复医学杂志

Objective:To explore the therapeutic effect of comprehensive rehabilitation nursing on patients with cere-bral infarction (CI)complicated type 2 diabetes mellitus (T2DM).Methods:According to number table,a total of 118 CI + T2DM patients were randomly and equally divided into comprehensive nursing group and routine nursing group.Comprehensive nursing group received comprehensive nursing based on routine nursing.The evaluating inde-xes included fasting blood glucose (FBG)concentration,limb muscle strength recovery and score of activity of daily living of two groups before hospitalization and discharge.Results:Compared with routine treatment group before discharge,there was significant reduction [(7.9±4.2)mmol/L vs.(5.9±2.8)mmol/L]in FBG concentration in comprehensive nursing group;limb muscle strength recovery of comprehensive nursing group was significantly better than that of routine nursing group (≥ grade III,upper limb 83.1% vs.64.4%,lower limb 76.3% vs.55.9%);percentage of patients with activity of daily living score > 60 scores (33.9%)in comprehensive nursing group was significantly higher than that of routine nursing group (15.2%),P <0.05 all. Conclusion:Comprehensive rehabili-tation nursing could promote the rehabilitation of physical function in patients with cerebral infarction complicated type 2 diabetes mellitus.

Mycotic Pharyngitis or Stomatitis in Patients after Tracheotomy:Their Correlative Factors / 中华医院感染学杂志

OBJECTIVE To understand the factors of mycotic pharyngitis or stomatitis for patients after tracheotomy,especially related to antibiotic.METHODS Seventy patients after tracheotomy were divided into two groups.In each group,the clinical data,risk factors and culture results of oral cavity were analyzed.RESULTS The time,kind,dose,way,and combined use of antibiotic were all related to mycotic pharyngitis or stomatitis.CONCLUSIONS The longer time,the complex kind,the larger dose,and the combined use of antibiotic are all delayed patients recovering.

Relationship Between Optimal Time Duration to Use Peripheral Vein Catheter and Bacterium Colonization in Summer Season / 中华医院感染学杂志

OBJECTIVE To study the relationship between how long to use peripheral vein catheter and bacterium colonization,then to make an approach to the optimal time duration of using peripheral vein catheter in summer season from infection prevention viewpoint.METHODS To divide the patients into four groups according to the different time duration of using peripheral vein catheter,and select 20 samples at random in every group.Make bacterial culture from skin(before sterilization,after sterilization and withdrawal of needles) and the catheter,and compare the colony number and strain distribution.RESULTS There was bacterium colonization on punctured skin after using peripheral vein catheter for a long time.The longer it was,the higher the positive rate was.After 96 hours there was bacterium colonization in vein catheter(10%) and after 120 hours positive rate was obviously higher(40%).CONCLUSIONS After using peripheral vein catheter for 96 hours in summer season,there is bacterium colonization in catheter,increasing the risk for infection,so the optimal time duration of using peripheral vein catheter in summer is 72-96 hours.

Drug Resistance to Respiratory Tract Infection of Patients after Tracheotomy / 中华医院感染学杂志

OBJECTIVE To acknowledge the important factor of bacterial infection of patients after tracheotomy. METHODS Thirty five patients′ lower respiratory tract sputa were sent to conduct bacterial culturing and determine their susceptibility to antibiotics and analyze results with statistics methods. RESULTS Total 8 main kinds of bacteria were isolated and drug resistance rate of Gram-positive cocci over 50 percent was erythromycin,that of(Gram-negative) bacilli included ampicillin,gentamicin,ciprofloxacin,and cefuroxime.The average time of antibiotics usage among 35 patients was(27?19.17) days and 12 kinds of antibiotics were used. CONCLUSIONS The time of usage of different antibiotics is an important factor to aggravate patients′ infection after tracheotomy.

DETECTION AND SIGNIFICANCE OF OCCULT NODAL MICROMETASTASIS IN COLORECTAL CARCINOMA / 肿瘤研究与临床

Objective To study the expression and significance of CEA in occult micrometastasis ofcolorectal carcinoma. Methods 287 cases without metastatic lymph nodes were observed in 54 cases ofcolorectal carcinoma by immunohistochemical (LSAB) method. Results 15 cases of lymph nodes werefound with occult micrometastasis in 8 patients. All micrometastasis was positive in detection of CEA.Conclusion Detection of CEA was conducive to detect micrometastasis of lymphnode. It is significant forDukes staging,prognostic determination and treatment.

Clinical Application of the Implantable Biventricular Pacing Cardioverter Defibrillator in 5 Patients / 昆明医科大学学报

Objective To analyze 5 patients suptered heart failure and ventricular tachycardia((VT).)Methods The Implantable Biventricular pacing cardioverter defibrillator was used to analyzed 5 patients(4 with primary dilated cardiomyopathy and 1 with coronary heart disease) suffered from heart failure and ventricular tachycardia(VT).Results 2 cases had history of syncope,4 patients were implanted INSYNC 7272 implantable cardioverter defibrillator(ICD) and 1 patient was implanted V-350 ICD.During the 1~14 month follow-up period,all the cardio functions were improved without VT or syncope.1 patient with VT and VF was detected and terminated by ICD.The patient was saved.Conclusions Implantable biventricular pacing cardioverter defibrillator is an effective approach to treat sudden cardiac death,mortal ventricular arrhythmia and heart failure.

Comparison of the Effect of Atrioventricular Nodal Ablation and Permanent Pacing Versus Atrioventricular Nodal Modification in Patients with Atrial Fibrillation / 昆明医科大学学报

Objective To evaluate the effect of atrioventricular nodal ablation and permanent pacing(AVNAP) or atrioventricular nodal modification(AVNM) in the treatment of atrial fibrillation(AF).Methods 32 patients with drug refractory AF were divided into AVNAP group(18) and AVNM group(14).The patients were measured the left ventricular ejection fraction(LVEF) by the echocardiogram,tested by the 6 minu te walking test(6-MWT),assessed by NYHA function classification and the Mos 36~item short from health surrey(SF~36) before or after the treatments respectively.Results Mean follow up is 14 months.In AVNAP group,LVEF was increased from(47?9)% to(57?6)%(P

Complication and Treatment of Undergoing Interventional Therapy for Cardiovascular Diseases / 昆明医科大学学报

Objective To observe the occurrence of the complication in patients under going interventional therapy for cardiovascular diseases and analyze its causes.Methods The data records of 1015 patients with interventional therapy for cardiovascular diseases in our hospital from October,2003 to October,2004(aged between 7 and 85 years,585 males and 430 females) were analyzed.Among them,520 patients received percutaneous transluminal coronary angioplasty and stent implantation,160 underwent radiofrequency catheter ablation,201 with permanent cardiac pacing,121 received blocking for congenital heart disease,10 received intraartery stent implantation and 29 with other interventional therapy.Results The complication occurred in 7.58% of 1015 patients.A higher incidence of hypotension and aberrant reflex has been reported(2.66% of all,mortality rate 0.39%).Conclusions Interventional therapy for cardiovascular diseases is an effective treatment with some risk.Serious pre-operational appraisal,enriching the experience of the doctors and strict post-operational monitoring may reduce the occurrence of the complication.

Clinical Research of Implantable Cardioverter Defibrillator / 昆明医科大学学报

Objective To report the clinical oberservation of 50 patients with implantable cardioverter defibrillator(ICD).Methods Observe the patients with ICD from May,1998 to Nov.2005.Results There were more than a thousand episodes of ventricular tachycardia ventricular fibrillation(VT/VF) detected and terminated by ICD devices.Conclusions ICD with tiered therapy function has high efficacy on the termination of ventricular tachyarrhythmias.It is important to follow up the patients and dynamically optimize the system of ICD.

Clinical Study on Radiofrequency Ablation for Complicated Arrhythmia Guided by Intracardiac Noncontact Catheter Mapping System / 昆明医科大学学报

Objective To evaluate the efficacy and superiority of intracardiac noncontact catheter mapping system for mapping and ablation of complicated arrhythmia.Methods 12 patients(9 males and 3 females,aged between 28 and 76 years) were included in this study.The characteristics of electrophysiological examination were as follows one was idiopathic ecotopic beats originated in right ventricular outflow tract(RVOT),two cases with idiopathic left ventricular tachycardia(ILVT),one atrial tachycardia(AT) of left atrium,two AT originated in right atrium,six atrial fibrillation(Afi) of left atrium.Among them,4 cases failed with conventional electrophysiological examination and radio frequency ablation(RFA),1 patient was reported recurrence.All patients were introduced 64 non-contact unipolar electrodes and ablation catheter to the same chamber via formal veins,reconstructed three-dimension geometry configuration,isopotential maps and analyzed the foci of reentrant activation and critical isthmus,then ablated the interested sites guided by noncontact mapping system.Results One idiopathic ectopic beat originated in RVOT near septum received segmental ablation.Two ILVT substrates located at left posterior-mid septum and left-posterior septum underwent segmental ablation.One AT of left atrium(LA) originated in right inferior pulmonary vein(RIPV) was given linear ablation between RIPV and mitral.One of the two AT from right atrium(RA) mapped the earliest activation in orifice of inferior cava vein and was given cricoid ablation;another AT originated in posterior wall of superior cava vein across crista terminal received linear lesion.One of the six Afi cases triggered by left atrial ectopic beats was given linear lesion between left superior pulmonary vein(LSPV) and right superior pulmonary vein(RSPV) during sinus rhythm;the other five cases mapped the reentrant origin site of Afi during persistent Afi rhythm received the catheter linear ablation,and the ablation lines respectively were created between RSPV and RIPV,LSPV and left inferior pulmonary vein(LIPV),LSPV and RSPV,LIPV and mitral isthmus.All patients succeeded and no obvious complication observed in operation and postoperation.None recurred during 2~26 month follow-up period.Conclusions It is safe and effective of intracardiac noncontact catheter mapping system for endocardium mapping and ablation of arrhythmia,especially for the clarity of electrophysiological mechanism of complicated arrhythmia and guiding RAF,which has potential clinical values.