1.Current status and prospects of laboratory diagnosis of clinical virus infection
Chinese Journal of Laboratory Medicine 2014;(10):728-731
The severe infectious diseases caused by virus are occurring with increasing frequency, which poses a rising global threat to human health and life.There are many kinds of viruses that cause a wide variety of viral diseases.The same kind of virus is able to cause different diseases, meanwhile a disease can be caused by different viruses.All these conditions make the relationship between viral pathogens and infection diseases complicated.At present, no effective laboratory detection methods for most of virus infectious diseases are developed.But the rapid development of molecular diagnostic technologies makes it possible for clinical laboratory to detect viral infection diseases rapidly and simultaneously.This review summarized the present and development perspectives of laboratory tests for the diagnosis of clinical virus infections, attempting to give some advices for laboratory diagnosis procedure of viral infections.
2.The expression of MMP-1, COX2 induced by IL-1? in human dental pulp cells
Qun LU ; Xuedong ZHOU ; Jun CAO
Journal of Practical Stomatology 1995;0(04):-
Objective:To study the effect of endogenous IL-1? on the expression of matrix metalloproteinases (MMPs) and COX2 in dental pulp cells. Methods:Human dental pulp cells were treated with human recombinant IL-1? at 1 nmol/L in serum-free medium for 18 h. Then the cells were collected and total RNA was isolated, MMPs and COX2 mRNA expression was assessed by semiquantitative RT-PCR. Results:IL-1? at 1 nmol/L induced the expression of COX2 and MMP-1 mRNA in human dental pulp cells. Conclusion:IL-1? may contribute to stimulating expression of MMPs and COX2 in the dental pulp during pulpitis.
3.Research Advancement of TOF-MS for Analysising Protein Components in Patients with Diabetic Nephropathy Urine
Ning ZHANG ; Xuedong LU ; Zhenglin WU
Journal of Modern Laboratory Medicine 2016;31(5):160-164
TOF-MS has been more and more widely used in clinical and scientific research due to its advantages such as fast and easily operated.Diabetic nephropathy is the most common complications of diabetes.But clinical existing detection meth-ods to some extent have some hysteresis.The applications and developments of TOF-MS coupled with other proteomics technologies provided new ideas and methods for early diagnosis and prognosis of diabetic nephropathy.In this paper,the lat-est research results in nearest years about the application of TOF-MS to diabetic nephropathy will be reviewed.
4.Study of expression level of acute lower respiratory tract Boka virus infectecd children with serum specific antibody
Yunqi ZHANG ; Qiong WANG ; Yinhui ZHANG ; Xuedong LU ; Taojun HE ;
International Journal of Laboratory Medicine 2015;(12):1660-1662
Objective To discuss the clinical value of the expression level of acute lower respiratory tract Boka virus (HBoV)in‐fectecd children whose detection serum specific antibody .Methods 904 cases of children with acute lower respiratory tract Boka vi‐rus infection hospitalized from March 2011 to July 2014 who were selected as study objects ,serum ,sputum ,bronchoalveolar lavage fluid HBoV DNA positive were as the gold standard for diagnosis of acute lower respiratory tract infection in HBoV ,the positive serum HBoV antibody of HBoV in children with acute lower respiratory tract infection was defined as the observation group ,serum HBoV antibody negative acute lower respiratory tract infection in children with HBoV was defined as the control group ,the correla‐tion between serum HBoV antibody and acute lower respiratory tract HBoV infection children whose clinical characteristics were analyzed .Results Serum HBoV antibody in the diagnosis of acute lower respiratory tract infection of HBoV whose sensitivity ,spe‐cificity ,positive predictive value ,and negative predictive value ,accuracy of diagnosis were separately 60 .32% 、90 .25% 、31 .67% 、96 .81% 、88 .16% .In the general data ,between the observation group and the control group in gender ,age ,hospitalization time , there were no significant differences(P>0 .05) .In the clinical manifestations ,nasal congestion and runny nose ,cough ,fever ,vomi‐ting and diarrhea ,shortness of breath ,breathing difficulties whose occur rates had no significant differences between the observation group and the control group(P>0 .05) ,the incidence of wheezing of the observation group was significantly higher than that of the control group ,the difference had statistical significance (P<0 .05) .The comparison of clinical diagnosis between the observation group and the control group had no significant difference(P>0 .05) .Conclusion Serum HBoV antibody is in favor of acute lower respiratory tract infection of HBoV in the diagnosis of exclusion ,and the serum HBoV antibody positive and acute lower respiratory tract infection of HBoV have a certain relationship in children with wheezing symptoms .
5.Quantitative Analysis of Lung HRCT in Asymptomatic Nonsmokers and the Relationship with Pulmonary Function Test Results
Xuedong YANG ; Xiaodong LU ; Wei ZHOU ; Shaoke LI
Journal of Practical Radiology 2001;0(10):-
Objective To study the changes with aging of lung density parameters on inspiratory and expiratory HRCT in asymptomatic nonsmokers,and the correlation with pulmonary function test(PFT).Methods Lung HRCT scans were performed at end inspiratory and end expiratory in 63 subjects,and PFT were performed in 32 of them.All the subjects were divided into 5 groups according to the ages.Lung HRCT quantitative parameters of each groups were measured,and the correlation with PFT results was assessed.Results In each age group,the mean lung density of expiratory lung HRCT and the overall lung density difference between inspiratory and expiratory HRCT was significant decrease(P
6.Comparison of five methods used for detection of Clostridium difficile infection
Ying HE ; Xuedong LU ; Haijing LI ; Shufang MENG ; Yiwei TANG
Chinese Journal of Laboratory Medicine 2010;33(12):1139-1144
Objective To evaluate five detection methods for the laboratory diagnosis of Clostridium difficile infection in the hospitals of USA, and explore a sensitive, specific, accuracy and rapid regimen for the early diagnosis of Clostridium difficile infection. Methods A total of 174 stool specimens submitted to the clinical microbiology laboratory for Clostridium difficile testing were separately tested by five methods including toxigenic culture (TGC), Premier Toxin A&B EIA( A/B-EIA), C. Diff Quick Chek Complete( DEIA), BD G eneOhm Cdiff assay(BD-PCR) and Laboratory-developed PCR(LD-PCR). The gold standard of TGC was used as a reference criterion, and the sensitivity, specificity, positive predictive value ( PPV )and negative predictive value (NPV) of A/B-EIA, D-EIA, BD-PCR and LD-PCR assays were determined. Results Among the 174 specimens studied, 24 were defined as true positives for Clostridium difficile infection by TGC assay, giving a positive rate of 13.8% (24/174). In comparison to the standard,the sensitivity, specificity, PPV and NPV were 62.5%, 99.3%, 93.8% and 94.3% for A/B-EIA;66.7%, 98.7%, 88.9% and 94.9% for D-EIA; 83.3%, 98.7%, 90.9% and 97.4% for BD-PCR;79.2%, 93.3%, 65.5% and 96.6% for LD-PCR. Among all tested specimens, 34 were positive by atleast one of five methods, and of which 15 were concordant by all five methods. The D-EIA results were divided into three groups depending on results of GDH and (or) toxins A/B: 18 were positive for both GDH and toxins A/B, 23 were positive for only GDH, and 133 were negative for both GDH and toxins A/B. Of 18 positive specimens by D-EIA assay, all were concordant with results of BD-PCR assay and 16 were agreement with results of TGC assay. Twenty-two of 24 positive specimens by TGC assay were included in 41 specimens that were positive for GDH. Among eight false negative specimens by D-EIA assay, four were differentiated as positive results by BD-PCR. According to the present study, the sensitivity, specificity,PPV and NPV of a two-step detection algorithm in combination with D-EIA and BD-PCR assays were 83.3%, 98.7%, 90.9% and 97.4%, respectively. Conclusions From the point of technological evaluation, BD-PCR is preferable. A two-step detection algorithm combining D-EIA with BD-PCR is proposed for the laboratory diagnosis of Clostridium difficile infection. This algorithm has demonstrated an excellent sensitivity and specificity, as well as decreased test turnaround time and test cost.
7.Effects of purified product from trapa manshurica fler on apoptosis and cell cycle progression of tumor cells in vitro
Zhe LU ; Shouliang GONG ; Fenglan NIU ; Xuedong WANG
Journal of Jilin University(Medicine Edition) 2006;0(01):-
Objective To observe the effects of 3,4,5-trihydroxybenzoic acid dimmer (TAD) in different doses on apoptosis and cell cycle progression of liver cancer SMMC-7721 cells in vitro and explore its possible mechanism of inhibiting tumor growth.Methods The experiment was divided into 0,3.125,6.250,12.500 and 25.000 ?g TAD groups. Apoptosis was observed with JEM-1200EX transmission electron microscope (TEM).The changes of apoptosis and cell cycle progression were measured with flow cytometry.Results The typical features of apoptosis were found in tumor cells,the nuclei were broken to pieces,mitochondrion cristae were disrupted and vacuoles were showed in nucleus and cytoplasm in 25.000 ?g TAD group observed under TEM.Meantime,the apoptotic percentages of the cells treated with 3.125—25.000 ?g TAD were increased significantly as compared with control group (P
8.Analysis of cultured and secreted antigens in Mycobacterium tuberculosis H_(37)Rv
Xuedong LU ; Yinhui ZHANG ; Xiaoyan ZHANG ; Laizhi YANG
Chinese Journal of Laboratory Medicine 2003;0(08):-
Objective To analyze the antigens of solid cultured germ, liquid cultured germ and secreted composition, looking for the main immunity antigens of Mycobacterium tuberculosis H37Rv.MethodsShowing the difference of the antigens of solid cultured germ, liquid cultured germ and secreted composition by SDS-PAGE, Western blotting, monoclonal and polyclonal antibody techniques.Results Antigens of the solid cultured germ were nearly accordance with the liquid cultured germ and they were difference with secreted proteins. The main immunity antigens of the cultured germ were 79 000, 54 000 ,38 000~50 000, 28 000 and the secreted proteins are 66 000, 55 000~64 000, 30 000~34 000, 24 000, 16 000.The antigens of 66 000, 55 000 and 16 000 were proved that they were secreted proteins by monoclonal antibodies.Conclusions This study demonstrated that the main immunity antigens of the cultured germ in Mycobacterium tuberculosis H37Rv were difference with the secreted one .This is beneficial to the cognition of H37Rv and looking for special antigens for diagnosis of tuberculosis .
9.Expression of recombinant protein VP2 of human bocavirus and detection by indirect ELISA
Yunqi ZHANG ; Taojun HE ; Xuedong LU ; Yinhui ZHANG
The Journal of Practical Medicine 2016;32(17):2803-2806
Objective By prokaryotic expression and purifying the human bocavirus recombinant protein VP2, to establish the indirect enzyme-linked immunoassay for detection of virus. Methods We amplified the human bocavirus recombinant protein VP2 gene fragments from WHL-1 template by PCR , and cloned into the expression vector pET28a, then conversed into the BL21 (DE3) and expressed the fusion protein detected by Western Blot detection , the obtained the antibody and detected the human bocavirus in serum in Guanghzhou area in healthy people. Results The Recombinant prokaryotic expression identified correct by double enzyme, and it could occur specific reaction with the virus positive serum. The best optimal antigen coating concentration were serum multiples and blocking BSA was 2 mg/mL , 1 ∶ 200 and 1%. The best working dilution of enzyme-labeled secondary antibody was 1 ∶ 4 000. The best working hours was 1h. This detection method had good specificity and reproducibility. The cut-off of the indirect ELISA method was 0.1 and the sensitivity and specificity of the developed ELISA method were 92% and 98% respectively. The coincidence rate of determination results by the developed kit and control kit was 97%. Conclusion The competitive ELISA established by prokaryotic expressing and purifying the human bocavirus protein VP2 protein , provides a basis in detecting the human bocavirus serum antibody.
10.Study on the Relationship between Patients with Hepatitis B Viral Loads and Immunoglobulin A,G,M and Complement C3,C4
Taojun HE ; Zhenglin WU ; Xiaoqiang ZHONG ; Shuping NIE ; Xuedong LU
Journal of Modern Laboratory Medicine 2015;(4):67-70
Objective To investigate the relationship between Hepatitis B patients with different viral loads and immunoglob-ulin A,G,M and complement C3,C4.Methods Firstly,followed by real-time fluorescence quantitative PCR detection 210 cases of hepatitis B patients with HBV-DNA levels,according to 10n copies/ml different viral load detection results,it was divided into 102 ~108 copies/ml of the experimental groups.Then the experimental groups and control group were simulta-neously detected in immunoglobulin A,G,M and complement C3,C4.Analysed the correlation between HBV loads and im-munoglobulin A,G,M and complement C3,C4.Results When the viral loads of hepatitis B patients were 105 ~108 copies/ml,the testing results of IgA,IgG and IgM were both increasing (U =12.43,10.96,6.42,P <0.01),while C3 and C4 were both decreasing (U =8.37,6.0,P <0.01).When the viral loads of hepatitis B patients were 102 ~ 104 copies/ml,only IgA and IgM were increasing (U =2.36,2.04,P <0.05),the other testing results had no statistical significance.Between the test of 7 experimental groups compared with each other,only 104 group and 105 group had significantly changed (IgA and IgM were increasing,C4 was decreasing,U =2.39,2.46,2.09,P <0.05,IgG was increasing,U = 3.25,P <0.01),but between other low viral loads or high viral loads were not significantly differences.Conclusion The different viral loads of hepatitis B patients could cause the different changes of immunoglobulin A,G,M and complement C3,C4,especially in the 4 groups from 105 to 108 copies/ml.Followed by increasing in viral loads,there were immunoglobulin A,G,M increasing and comple-ment C3,C4 decreasing,and also serious impaction on the immune function of organism.There was a positive correlation be-tween viral loads in vivo and immune damages,correlation coefficient (γ =0.967,P <0.01).When the viral loads from 104 to 105 copies/ml,the testing results had changed significantly.It suggest that should control viral loads under 104 copies/ml in the hepatitis B antiviral treatment,so the effect of immune function damage will be the minimum.