3.Construction of ICAT interference lentivirus expression vector and establishment of stable HL 60 cell lin
Jinshu WANG ; Weijia WANG ; Ting WANG ; Yan ZHANG
International Journal of Laboratory Medicine 2016;37(7):875-877
Objective To construct the ICAT gene interference lentivirus expression vector targeting and to establish stable transfected cell line HL60 .Methods The interference sequence targeted at human ICAT gene was designed and synthesized ,after annealing ,which was connected to PGLV3 interference vector and with PG‐p1‐VSVG ,PG‐p1‐REV ,PG‐p1‐RRE were co‐transfected into 293T cells The lentivirus particles were packaged and generated .The virus titer was detected .HL60 cells were transfected for establishing the stable cell line ;RT‐PCR and Western Blot techniques were used to detect ICAT gene and protein expression in sta‐ble HL60 cells ,then the results were compared with those in the control group .Results The lentivirus expression vector targeted at ICAT was successfully constructed and the virus titer was 2 × 108 U/mL .Stable transfected HL60 cell line was established .The effective interference verification revealed that shICAT could significantly reduce the mRNA and protein level of ICAT ( P<0 .001) .Conclusion The shRNA lentiviral expression vector of ICAT gene is successfully constructed and the HL 60 cell line stably interfering ICAT expression is established .
4.Diagnostic Value of Detecting the Serum and Urine Procalcitonin(PCT) for the Urinary System Infection
Haizhong YAN ; Weijia WANG ; Yuejing MU ; Quanzhong XU
Journal of Modern Laboratory Medicine 2017;32(4):116-118
Objective To explore diagnostic value of the serum and urine procalcitonin (PCT) detecting in the urinary system infection.Methods The serum and urine PCT levels in 45 urinary system infection patients with clear pathological diagnosis (exclude other system infections) who were outpatiented or hospitalized in the People's Hospital of Zhongshan between March and November 2016 (including 21 cases of upper urinary tract infection and 24 cases of lower urinary tract infection) and 35 healthy adults who went through physical examinations at the hospital during the same period,were measured using electrochemiluminescence immunoassay (ECLIA) on Cobase 601 Immunoassay Analyzer and analyzed to compare the differences of PCT levels in the three groups.Results The urine PCT level in upper urinary tract infection group was 0.243± 0.123 ng/ml.It was significantly lower than lower urinary tract infection group (0.486±0.232 ng/ml,t=4.11,P=0.000) and control group (0.454± 0.253 ng/ml,t=3.96,P=0.000).The serum PCT level in upper urinary tract infection group was 0.062±0.014 ng/ml.It was obviously higher than that in lower urinary tract infection group (0.043±0.020 ng/ml,t=3.56,P=0.01) and control group (0.032±0.013 ng/ml,t=7.38,P=0.000).In all groups,the urine PCT levels were significantly higher than their serum PCT levels (t =9.48,9.12,6.79,P< 0.01),and significant differences were observed in them.The sensitivity,specificity,positive predictive value and negative predictive value of serum PCT for diagnosing upper urinary tract infections were 81.5%,84.2%,80.6% and 85.6% respectively,and the urine PCT were 86.4%,80.7%,88.4 % and 83.1 % respectively.Conclusion Detection of serum and urine PCT has important accessory diagnostic value for identifying upper and lower urinary tract infections.
5.A preliminary study on Norovirus variant for GⅡ.4 in Guiyang regions
Yan YAN ; Jun GUO ; Weijia JIANG ; Kecheng TIAN ; Shijun LI ; Jing HU ; Guangpeng TANG ; Dingming WANG
Chinese Journal of Microbiology and Immunology 2012;32(7):606-609
Objective To study the composition of variant and point mutations of Norovirus GⅡ.4 in Guiyang regions.Methods From June to November 2010,cases information and fecal specimens were collected from guard-hospitals in Guiyang regions,who had caught the acute-gastroenteritis.Noroviruses in specimens were detected by a real-time reverse transcription polymerase chain reaction(real-time RT-PCR),and then partial genotyped norovirus-positive clinical samples (in random) were cloned and sequenced in VP1 gene code.Furthermore,the gene sequences were compared with the published variants at home and abroad of norovirus(GⅡ.4),including the phylogenetic analyses of genomes and variation of amino acids within individual sites.Results Those 267 specimens were GⅡ-norovirus-positive(62.68%) in 426 clinical samples.There were nine GⅡ.4-norovirus-positive VP1 gene-sequences available,and two subtype-norovirus variants (GⅡ.4 2008a and G Ⅱ.4 2008b variant) were epidemic in 2010,Guizhou province.The homology between and in subgroups were 95.90%-96.72% and 99.45%-100%.Two amino acids within individual sites were apt to mutate.Conclusion Norovirus GⅡ genotype were predominant in summer and fall acute gastroenteritis in 2010 for Guiyang regions,and the variants were diversity.
6.Analysis of factors influenceing patients' family to make decision to reject resuscitation to the patients
Jike XUE ; Shouquan CHEN ; Zhangping LI ; Huiping LI ; Weijia HUANG ; Junyan CHENG ; Jie ZHANG ; Ping YAN
Chinese Journal of Emergency Medicine 2009;18(11):1211-1214
Objective To study the factors influenceing patients' family members to make own relative fac-tors patients' families making decision on refusal of cardiopulmonary resuscitation (CPR) to the critical patients. Method Data were registered based on Utstein Style of 522 patients aged over 15 years, who subjected to in-hos-pital cardiac arrest(CA) in Department of Emergency of The First Affiliated Hospital of Wenzhou Medical College from January 2005 to December 2008. A total of 157 patients' family made refusal decision among the 522 pa-tients, who belonged to the refusal group, and others belonged to the attempt resuscitation group. The associated factors included age, sex, marriage, household register, cause of CA, underlying diseases, capability of activity before CA, life supported with mechanical ventilation, and administration of pressor agents. The refusal decisions were evaluated by using univariate Logistical regression analysis, and then the statistical significant variables were analyzed by using muhivanate Logistical regression analysis. Results Age, household register, cause of CA(car-diac or traumatic),stroke, sudden death, cancer, capability of activity before CA, life supported with mechanical ventilation,and administration of pressor agents were the important factors of making refusal decision (P < 0.01), but sexes or marriage was insignificant related to the refusal decision (P > 0. O5). The independent risk factors re-lated to refusal decision were age (P = 0.034),cancer (P = 0.006),stroke (P = 0.003), and life supported with mechanical ventilation (P = 0.000) in multivariate Logistical regression analysis, but the protective factors were sudden death (P =0.000),cardiac CA (P =0.020) and traumatic CA(P =0.000). Conclusions Age over 60 years, cancer, stroke, and life suppoted with assisted ventilation before CA were factors associated with re-fusal decision making, yet sudden death, cardiac CA and traumatic CA were factors of accepting CPR.
7.A noninvasive and continuous method for blood pressure measurement using pulse wave.
Haijiao LU ; Zhuangzhi YAN ; Weijia LU
Chinese Journal of Medical Instrumentation 2011;35(3):169-173
This paper presents a method of noninvasive and continuous measurement of blood pressure by pulse wave. The method is realized by extracting the characteristic parameters of human brachial artery pulse wave and developing the stepwise regression equation. The experiment results showed that blood pressure measured by this method was well correlated with which measured by mercury sphygmomanometer. The mean difference of blood pressure was smaller than 3 mmHg, and the standard deviation was smaller than 5 mmHg.
Blood Pressure
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physiology
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Blood Pressure Determination
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methods
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Blood Pressure Monitors
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Humans
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Pulse
8.Evaluation of NT-proBNP assay performance on electro-chemiluminescence immunoassay system
Xiuming ZHANG ; Dongmei WEN ; Weijia WANG ; Fei LI ; Lijuan KAN ; Haizhong YAN ; Minghuan SUO ; Nengliang OUYANG
Chinese Journal of Laboratory Medicine 2011;34(12):1152-1157
ObjectiveTo evaluate analytical performance of NT-proBNP on Electro-Chemiluminescence Immunoassay system.MethodsThe precision,accuracy,limit of blank ( LoB ),limit of detection (LoD),functional sensitivity (FS),analytical measure range (AMR),maximal dilution rate,clinical reportable range(CRR) and the analytical anti-interference ability of NT-proBNP were evaluated according to EP documents issued by CLSI and related references.The analytical performance data were compared to quality standards declared by the manusfacturers.According to CLSI C28-A2,80 healthy volunteers,aged from 18 to 74, were chosen and divided into 4 groups on average for biological reference intervals verification.Results The within-run CV and total CV were 1.1% -2.2%and 1.5% -2.9% respectively.The deviations from controls distributed by National Center for Clinical Laboratory and affiliated calibrators were 2.7% -5.9% and 2.7% -7.5%,respectively.The results of LoB,LoD and FS were 2.5,7.8 and 8.8 pg/ml,respectively.AMR was 8 -35 126 pg/ml,and the most suitable dilution rate was 1∶ 2,so the CRR was 9 -70 252 pg/ml.428 μmol/L bilirubin,2 g/L haematoglobin and 2 200 FIU chyle didn't interfere with the NT-proBNP assay.Moreover,almost all the data from different age groups were in the range of biological reference intervals declared by the manusfacturers, except one test data (167 pg/ml).Conclusions The analyticalperformance of NT-proBNP analyzed on Roche Cobas E601electrochemiluminescence immunoassay systemisconsistentwiththestandarlswhichmanufacturershas proclaimed.The establishment of LoD,FS,maximal dilution and CRR for NT-proBNP assay could provide the quality assurance for clinical use and the biological reference intervals declared by manusfacturers could meet the clinical needs.
9.Design and implementation of EMR information system in hospitals.
Weijia LU ; Zhuangzhi YAN ; Dengfu YAO
Chinese Journal of Medical Instrumentation 2011;35(6):437-441
With the electronic medical records information system as the core of hospital information platform design, this paper introduces the design of the system which is structured for electronic medical records, and the advantage and effectiveness is also introduced. From the aspects of the concept, system framework, data integration, right frame and safety control, identity index and a clinical pathway, etc, the paper introduces the basic idea and process of the hospital information platform design, with the hospital recording electronic medical records as the core.
Electronic Health Records
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Hospital Information Systems
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Medical Records Systems, Computerized
10.Effect of exogenous phosphocreatine on cardiomycytic apoptosis and expression of Bcl-2 and Bax after cardiopulmonary resuscitation in rats
Ping YAN ; Shouquan CHEN ; Zhangping LI ; Jie ZHANG ; Jike XUE ; Wantie WANG ; Weijia HUANG ; Junyan CHENG ; Huiping LI
Chinese Journal of Emergency Medicine 2011;20(7):698-702
Objective To observe the changes of cell apoptosis and levels of Bcl-2 and Bax protein in myocardium after cardiopulmonary resuscitation (CPR) in rats and to study the protective effects of different doses of exogenous phosphocreatine (creatine phosphate, CP) on cell apoptosis. Methods A total of 32 male adult SD rats were randomly divided into 4 groups, namely control group ( group A), CPR group (group B), low dose CP group (group C, phosphocreatine 0. 5 g/kg given at beginning of CPR and 1.0 g/ kg 2 hour after CPR) and high dose CP group ( group D, phosphocreatine 1.0 g/kg at beginning of CPR and 2. 0 g/kg 2 hours after CPR) . Cardiac arrest was induced by asphyxiation and CPR was started 7 min after asphyxiation it groups B, C and D. Myocardium samples were taken 24 hours after CPR for detecting myocardium cell apoptosis by TUNEL method. The levels of Bcl-2 and Bax protein were measured by using immunohistochemistry. Experimental data were processed with variance analysis in SPSS package. Results Compared with group A, myocardium cell apoptosis index (AI), and the levels of Bcl-2 and Bax proteinincreased significantly in groups B, C and D (P <0. 01 ), and Bcl-2/Bax ratio decreased significantly (P <0. 01 ) . Compared with group B, myocardium cell AI and levels of Bcl-2 and Bax protein decreased significantly in groups C and D ( P < 0. 01 ), and Bcl-2/Bax ratio increased significantly ( P < 0. 01 ) .Compared with group C, myocardium cell AI and levels of Bcl-2 and Bax decreased significantly in group D (P < 0. 05 ), and Bcl-2/Bax ratio increased significantly ( P < 0. 05 ) . Conclusion Exogenous phosphocreatine, especially inlarge dose, could inhibit apoptosis of myocardium cells and alleviate myocardium injury after CPR in rats.