1.A Comparative Study on Four Methods in the Diagnosis of Tuberculosis
Journal of Modern Laboratory Medicine 2016;(1):134-137
Objective To evaluate the value of four methods in diagnosis of pulmonary tuberculosis,including T-SPOT,fluo-rescent PCR,anti-TB antibody test,and acid-fast staining.Methods Retrospective analysis of 530 cases between January 2012 and December 2014 who had taken four methods,and calculated the sensitivity specificity,positive predictive value,neg-ative predictive value,Kappa value,Youden index,positive likelihood ratio,negative likelihood ratio,Pairedχ2 test.Consider clinical diagnosis as the gold standard.Results The sensitivity,negative predictive value,Youden index of T-SPOT were 95.90%,97.29%,0.82,respectively,and all of these were the highest.The negative likelihood ratio of T-SPOT was 0.05, which was the lowest.Misdiagnosis rate of PCR was 87.18%,which was the highest.Positive likelihood of anti-TB antibody test was the lowest,6.48,while other indicators were no advantage.The specificity,positive predictive value and positive likelihood ratio of acid-fast staining were 99.70%,98.90%,153.83,respectively,and the three of these were highest.Pair-wise comparison between the four methods were significantly different.Conclusion The T-SPOT and acid-fast staining can be used as important methods,and the anti-TB antibody can provide result quickly,and PCR method is more suitable for ex-amination of sterile body fluids.
2.Different activities of antimicrobial combinations against muitidrug resistant Acinetobacter baumannii in vitro
Chinese Journal of Microbiology and Immunology 2011;31(10):898-902
Objective To evaluate the activities of 18 pairs of antimicrobials combinations against non - duplicate clinical isolates of multidrug resistant Acinetobacter baumannii (MDRAB) in vitro.Methods Collect isolates of Acinetobacter baumannii from different patients from October 2009 to May 2010,which were isolated in Clinical Laboratory Center of Beijing Friendship Hospital,Capital Medical University.Use broth microdilution method to detect MIC of mono-antimicrobial,and checkerboard broth microdilution method to detect combinatied MIC,and calculate fractional inhibitory concentration (FIC) index to determine drug combinations effects.When the performance of the same drug combinations conflicted,appropriate strains were selected for screening of drug-resistant mechanisms by polymerase chain reaction( PCR),including efflux pump genes.Results In tests in vitro,rifampicin and polymyxin B,imipenem and gentamicin,cefepime and levofloxacin showed synergy at high proportion,68.1%,45.5%,40.9%,respectively.Minocycline and rifampicin,ampicillin/sulbactam and tobramycin.Ceftazidime and ciprofloxacin showed additive effect at high proportion,81.8%,68.2%,68.2%,respectively.There were several combinations which appeared the opposite effects to tested strains.Strains No.19 corresponding reaction was synergy and No.21,No.26 corresponding reactions were antagonism.The three strains above were selected for screening resistant mechanisms.The difference is that genotypes of adeS were negative in No.19 and positive in No.21 and No.26.Conclusion Rifampicin and polymyxin B combination showed synergy against the MDRAB in vitro,which can be considered as the treatment choice for critical infections caused by MDRAB.Imipenem and gentamicin,cefepime and levofloxacin also showed synergy in vitro,but in some isolates showed antagonism.This phenomenon may be due to the gene adeS activated by certain antibiotics,and the activated adeS drived efflux pump express or overexpress,which made the drugs in bacterial cells pumped out,causing antagonistic effect.The individual differences in strains should be considered when clinic strain apply these two combinations above.
3.Combined antimicrobial susceptibility test against pan-drug-resistant Acinetobacter baumannii with E-test and microdilution checkerboard assay
Chinese Journal of Microbiology and Immunology 2013;(2):144-147
Objective To evaluate the activity of antibiotics against pan-drug-resistant (PDR) Acinetobacter baumannii by combination antimicrobial susceptibility test in viro with epsilometric methods (Etest method) and microdilution checkerboard (CB method),and to detect a good correlation between timekill curve with the above mentioned two assays.Methods Thirty-one clinical isolates of PDR Acinetobacter baumannii were selected for mono and combination antimicrobial susceptibility test in vitro by E-test and CB method,then a comparison was conducted between the test results and the time-kill curve.Mono drugs involved tigecycline,colistin,imipenem and amikacin,and combinations involved two of drugs above,and three drugs involved imipenem/tigecycline,plus amikacin combination.Results Synergistic effect was detected in imipenem plus colistin and tigecycline plus imipenem combination.A high comparability was revealed between the E-test method with antimicrobial drugs added into the culture medium and the time-kill curves.Synergy in the combination of imipenem/tigecycline,plus amikacin was detected by the CB method and time-kill curves.Conclusion The results showed that the effect of specific combination of antibiotics against PDR Acinetobacter baumannii could be predicted by testing their synergistic effect with combination antimicrobial susceptibility test.
4.Advances in the study of Mycobacterium tuberculosis protein phosphatase and its inhibitors.
Tieshan TENG ; Honghai WANG ; Jianping XIE
Acta Pharmaceutica Sinica 2011;46(12):1420-8
Reversible protein phosphorylation regulates multiple biochemical events. Mycobacterium tuberculosis phosphatases play important roles in regulating the pathogen physiology and interference of host signaling. They are also involved in the evasion of host immune response and blockage of the phagosome-lysosome fusion. Selective inhibition of phosphatase represents an ideal new avenue of anti-tuberculosis drug design. In this paper, we update the progresses about the regulation network of Mycobacterium tuberculosis phosphatases including MptpA, MptpB, MstP, SapM and their inhibitors. These serve as the basis for further antituberculosis drug target.
5.Quantitative analysis on EB virus DNA in peripheral white blood cell,plasma and serum in patients with hematologic diseases
Kedi WANG ; Zhi LV ; Tieshan WANG ; Jianrong SU
International Journal of Laboratory Medicine 2014;(16):2161-2162
Objective To compare the load of EB virus in peripheral white blood cell,plasma and serum in the patients with he-matologic diseases,and to discuss the feasibility of detecting EB virus load by using plasma or serum instead of peripheral white blood cell.Methods Venous blood of 125 patients with hematologic diseases were collected,and peripheral white blood cell,plasma and serum were isolated.The real-time quantitative PCR was used to detect the virus load in three kinds samples with the EB virus load in peripheral blood cell as the gold standard.Results Compared to peripheral white blood,the EB virus load in plasma and ser-um showed no statistical difference(P >0.05).Conclusion Using plasma or serum instead of peripheral white blood cell for con-ducting the quantitative detection of the load of EB virus will be a reliable method.
6.Three-stage swallowing rehabilitation for stroke patients with dysphagia
Ying GU ; Tieshan LI ; Hong CHEN ; Jinling WANG ; Yongtao LV
Chinese Journal of Physical Medicine and Rehabilitation 2009;31(4):261-263
Objective To explore the effects of three- stage swallowing rehabilitation on the swallowing a-bility of stroke patients with dysphagia. Methods 60 stroke patients were divided into primary cerebral infarction and primary cerebral hemorrhage groups, then further divided into treated and control groups randomly. All groups were given the same routine internal medicine treatment. Patients in the treated group were given three stage swallo-wing rehabilitation training additionally. All patients were assessed using Caiteng's Grading Method at the outset and at the end of the 2nd week, the Ist month and the 2nd month. Results Swallowing function scores in the treated groups were higher than those in the control groups at every stage (P≤0.05). The treated groups' scores also im-proved more quickly than those in the control groups. Conclusion Three stage swallowing rehabilitation can signifi-cantly improve stroke patients' swallowing function.
7.Expression of Fascin-1 Protein in Colorectal Adenocarcinoma and Relation with Its Clinicopathologic Characteristics
Hongwei LI ; Tieshan WANG ; Yong HUANG ; Yanhong LI ; Haixia HU
Chinese Journal of Bases and Clinics in General Surgery 2003;0(04):-
Objective To investigate the expression of Fascin-1 protein in colorectal adenocarcinoma,and the relationship with its clinicopathologic features.Methods The expressions of Fascin-1 protein in colorectal adenocarcinoma tissues of 60 cases,colorectal adenoma tissues of 30 cases and normal mucosa tissues (4 cm distance to neoplasm) of 30 cases were detected by Microwave-EliVisionTM immunohistochemistry method,and the relationship between the expression of Fascin-1 protein in colorectal adenocarcinoma tissues and its clinicopathologic characteristics was analyzed.Results The expression of Fascin-1 protein was located in cytoplasm.The positive expression rates of Facsin-1 protein were 3.3% (1/30),30.0% (9/30) and 53.3% (32/60) in normal mucosa tissues,colorectal adenoma tissues and colorectal adenocarcinoma tissues,respectively.The expression of Fascin-1 was gradually increased in these three tissues,and there was statistical difference among the three tissues (P0.05).Conclusion The high expression of Fascin-1 protein is correlated to high invasion ability and lymph node metastasis,which can play as a sensitive index in predicting the invasion and metastasis of colorectal adenocarcinoma.
8.Establishment of doctor-oriented j ob evaluation factor system for general hospitals and survey analyses on its reliability and validity
Yang WANG ; Dan ZHANG ; Shuqiang XU ; Tieshan ZHANG
Journal of Jilin University(Medicine Edition) 2014;(6):1308-1313
Objective To investigate the highly-recognized job evaluation factors among the doctors from tertiary general hospitals and to establish a doctor-oriented job evaluation factor system for general hospitals followed with the reliability and validity analyses.Methods A questionnaire about the highly-recognized 23 job evaluation factors was launched among 791 doctors by the stratified sampling from six tertiary general hospitals in Beijing,and the factors were from a job evaluation model of tertiary general hospitals.Initial factor solutions were obtained by the principal components analysis of all evaluation factors and the main factors whose eigenvalues were over one were extracted as evaluation dimensions. Factor loadings were attained through Varimax and the factor whose factor loading was lower than 0.5 was eliminated.A reliability analysis by calculating Cronbachαcoefficient and a validity analysis with structural equation modeling were sequentially conducted on the job evaluation factor system.Results A doctor-oriented job evaluation factor system for general hospitals including 22 factors subject to 3 dimensions was established while the factor of job relevance was removed. The internal consistency coefficients of the dimensions were greater than 0.8 according to the calculation of Cronbachα, which showed a nice reliability. Several main indexes for evaluating the model fitting were all close to 0.9,which indicated a fair structural validity. Conclusion A doctor-oriented job evaluation factor system for general hospitals is established with fair reliability and validity,which could provide more references to the job evaluation of the doctors in tertiary general hospitals.
9.PCR-induced Modification of C Terminus of HPV-16 E7 and Expression of Mutational E7 in Eukaryotic Cells
Yagang ZUO ; Jiabi WANG ; Fang LIU ; Yan YAN ; Tieshan ZHU ; Donglai MA ; Baoxi WANG
Chinese Journal of Dermatology 1994;0(02):-
Objective To induce the mutation of HPV-16 E7 in two zinc-binding motifs near the C terminus by polymerase chain reaction (PCR) and evaluate the effect of this mutation on the antigen-specific immunity of HPV-16 E7. Methods HPV-16 E7 fragment was amplified by PCR and cloned into pGEM-3zf vector. Two site mutations at 58 and 91 animo acid sites in the open reading frame of HPV-16 E7 were induced by PCR, and then the molecular clones of HPV-16 E7 wild type (pcDNA3.1/E7) and mutant (pcDNA3.1/ME7) were successfully reconstructed. Western blot and immunofluorescence were used to detect the expression of E7 protein. Results Intracellular fluorescence signals were observed in the cells transfected with pcDNA3.1/E7 and pcDNA3.1/ME7 24 hours after transfection, but the signals in the cells transfected with pcDNA3.1/ME7 disappeared 48 hours after tansfection. Twenty-four and 48 hours after transfection with pcDNA3.1/ME7, E7 protein was not detected by Western blot. Conclusions The stability of HPV-16 E7 protein is reduced by mutations (C58G, C91G) near two zinc-binding motifs. It is suggested that the zinc-binding motifs near the C terminus of HPV-16 E7 may be important for maintaining the stability of E7 protein.
10.Botulinum toxin injection can reduce arthritis pain by inhibiting the expression of calcitonin gene-related peptide in the dorsal root ganglia
Chenglei FAN ; Xiao CHU ; Kaile WANG ; Lin WANG ; Zhenyuan NIU ; Tieshan LI
Chinese Journal of Physical Medicine and Rehabilitation 2016;38(9):652-658
Objective To explore the analgesic effect of intra-articular botulinum neurotoxin type A (BoNTA) injection in rats with adjuvant-arthritis pain,to quantify the expression of calcitonin gene-related peptide (CGRP) in the dorsal root ganglia (DRG) associated with arthritis pain,and to investigate the retrograde axonal transport of BoNT-A into the DRG after peripheral injection.Methods Ninety Sprague-Dawley rats were randomly divided into groups A,B,C,D and E,each of 18.A murine model of adjuvant-arthritis pain was established by injecting 50 μL of complete Freund's adjuvant into the left ankle in all the mice except those in group A.The control group A was treated with intra-articular injection of 50 μL of saline solution.Three weeks later,groups A and B were treated with a 20 μL intra-articular saline injection,while groups C,D and E received an intra-articular injection of BoNT-A at 1 U/20 μL,3 U/20 μL or 10 U/20 μL respectively.Pain threshold and muscle strength were graded before and 1,5,15 and 21 days after the modelling,as well as at 1,3,5 and 14 days after the BoNT-A treatments.Protein expression and the CGRP-positive cell number were observed,as well as any BoNT-A-cleaved synaptosomal-associated 25 kDa protein (cl-SNAP-25) in the DRG using Western blotting and immunofluorescence.Results Compared with group A,there was a significant decrease in the average mechanical withdrawal threshold and muscle strength and a significant increase in the protein expression and the CGRP-positive cell number in the other 4 groups.Compared with group B,the mechanical withdrawal threshold had increased significantly more in groups D and E at 5 days after the BoNT-A injection and in group C at 14 days after the treatment.Compared with group B,the protein expression and the number of CGRP-positive cells were significantly lower in groups D and E at 3 days after the BoNT-A injection.The decrease in group C was significant after 14 days.No significant differences were found between groups D and E in any measurement at any time point.There was no significant difference among groups B,C and D in terms of muscle strength.Five days after the BoNT-A injection,significantly decreased muscle strength was observed in group E.In addition,BoNT-A cleaved-SNAP-25 was detected in the DRG.Conclusion BoNT-A can reduce arthritis pain through inhibiting the expression of CGRP in the DRG.Its analgesic effect has a dose response.A peripheral injection of BoNT-A can arrive at the DRG through retrograde axonal transport.