1.The significance of intensifying the standardization and regularization for antimicrobial susceptibility testing in clinical microbiology laboratory
Chinese Journal of Laboratory Medicine 2001;0(05):-
Standardization and regularization for antimicrobial susceptibility testing in clinical microbiology laboratory is obvious and vital to ensure reliable and reproducible results to guide early and rapid therapy for clinical physicians.Clinical microbiology laboratory should play an important role in antimicrobial therapy.
2.Endoscopic Ureteral Excision Using Holmium Laser in the Treatment of Ureterostenosis:Report of 52 Cases
Lin YUAN ; Xiaojian GU ; Qingyi ZHU
Chinese Journal of Minimally Invasive Surgery 2005;0(10):-
Objective To discuss the techniques and efficacy of Ho:YAG laser incision by using uteroscopy for ureterostenosis.Methods From July 2004 to April 2007,52 patients with ureterostenosis received ureteral incision by using Ho:YAG laser under a endoscope.Two double pigtail stents(F5 or F6) were placed in the ureters after the operation and left indwelling for 8 to 12 weeks.Ultrasonography and excretion urography were performed 3 to 6 months after extubation.Results Follow-up was available for 3 to 24 months(mean,17 months) in 46 patients,of which 40(87%) were cured after the treatment.In the cured patients,hydronephrosis,ureteral dilation,and ureterostenosis were improved,and the pain in the kidney region was relieved;none of them showed signs of infection.In the other 6 patients,4 were improved after the treatment(no deterioration of the symptoms of hydronephrosis,ureteral dilation,and pain in the kidney region,and no infection);and 2 failed(the symptoms of hydronephrosis and ureteral dilation deteriorated,and pain in the kidney region and infection were developed).Conclusion Endoscopic ureteral excision using holmium laser combined with indwelling of two double pigtail stents is effective and safe for ureterostenosis.
3.Compared research of selective culture media for Legionella and clinical practicability
Zhaohui HU ; Qingyi ZHU ; Benrong LIU
International Journal of Laboratory Medicine 2006;0(03):-
Objective To develop a selective isolation media which has a high isolation ratio for Legionella.Methods We compared the growth of Legionella pneumonia on BCYE? plate (with DGVP), BCYE?(with GVPC) and BCYE? (with CCCV) by colony count and compared with BCYE? plate. The comparison of isolation ratio of Legionella pneumonia from air-conditions cooling water with 3 kinds of BCYE? plate containing different anti-reagents was performed.Results The results of colony count on BCYE? plate (with DGVP) and BCYE?(with GVPC) was identical with that on BCYE? plate at same concentration. But the result of colony count on BCYE? plate (with CCCV) was half of the former 2 kinds of plate. 5 strains of Legionella pneumonia isolated from 9 central air-condition cooling water were used by BCYE? plate (with DGVP) and BCYE? (with GVPC), while only 2 strains of Legionella pneumonia were isolated by BCYE? plate (with CCCV). Legionella pneumonia wasn′t detected by BCYE? plate.Conclusion BCYE? plate with antibiotics system of CCCV showed obvious suppression to Legionella pneumonia, and so that there was a low isolation ratio to Legionella pneumonia. But BCYE? plate with antibiotics system of DGVP and GVPC had a few suppression and showed a high isolatio ratio for Legionella pneumonia. Moreover, BCYE? plate with antibiotics system of DGVP and GVPC can effectively inhibit non-Legionella and increase isolation ratio of Legionella. They should be the first selection media for Legionella from environment and clinical samples.
4.Real-time fluorescence polymerase chain reaction for detecting Legionella in sputum specimens of patients with pulmonary infection
Lei ZHU ; Chunru QI ; Xianghong ZHOU ; Zhenxing ZHANG ; Qingyi ZHU
International Journal of Laboratory Medicine 2015;(18):2674-2676
Objective To establish a real‐time fluorescent polymerase chain reaction and detect 16S rRNA gene of Legionella strains isolated from sputum specimens of patients with pulmonary infection by using this method .Methods 16s rRNA gene of Le‐gionella was used to design primers and probes .The reaction system and reaction conditions were optimized and the specificity ,sen‐sitivity and repeatability of this method were verified by detecting Legionella pneumophila ,non‐Legionella pneumophila and other bacteria .A total of 557 sputum specimens of patients with pulmonary infection were detected ,and PCR‐digestion identification method was carried out as control .Otherwise ,sequences of 16S rRNA were verified in patients with positive detection results .Re‐sults The results showed that all reference strains of Legionella were positive ,while all of other bacteria were negative ,and the sensitivity was 102 CFU/mL .Among sputum specimens collected from 577 cases of patients with pulmonary infection ,the positive rate of Legionella detected by using real‐time fluorescent PCR and PCR‐digestion identification method was 23 .1% and 19 .9% re‐spectively ,while the positive rate was 17 .2% by verifying the sequences of 16s rRNA .There were no statistically significant differ‐ences of positive rate among the three methods(P>0 .05) .Conclusion The real‐time fluorescent PCR is fast and convenient in de‐tection of Legionella strains isolated from sputum specimens of patients ,which could be an assisted method for clinically diagnosing Legionella infection .
5.Identification and characterization of 10 Francisella philomiragia strains
Lei ZHANG ; Daning YE ; Yan ZHU ; Haiyun CHAI ; Qingyi ZHU ; Cha CHEN ; Pinghua QU
Chinese Journal of Clinical Laboratory Science 2017;35(4):271-276
Objectives To identify and characterize 10 strains of Francisella philomiragia-like organisms isolated from blood samples and environmental water.Methods The 10 clinical and environmental isolates were identified by traditional morphological examination and biochemical characterization,matrix-assisted laser desorption/ionization time of flight(MALDI-TOF) mass spectrometry(MS) systems and sequencing based on 16S rRNA gene.The minimum inhibitory concentrations were tested by E-test methods.Results All the 10 isolates were gram-negative coccobacilli appearing tiny and faint counterstain of safranin,negative for urease,nitrate reduction and X and/or V factor requirement,but positive for oxidase and catalase.The isolates grew rapidly in sheep blood agar,chocolate agar and BCYE plate forming white opaque,colorless transparent or gray smooth colonies with about 2-mm diameters,but did not grow in M-H agar and MacConkey agar.The sequencing for 16S rRNA gene indicated that the 10 isolates shared more than 99.6% similarity to Francisella philomiragia,and fell into the same clusters of Francisella philomiragia on phylogenetic tree.The MALDI-TOF MS analysis also showed the typical peaks with 6 153 m/z,5 180 m/z,7 757 m/z and 9 392 m/z which were similar to Francisella philomiragia ATCC 25015.However,they may be misidentified to be Sphingomonas paucimobilis by using Vitek 2 GN cards,Neisseria cinerea by using Vitek 2 NH cards,Myroides odoratimimus by using API 20NE strips and Haemophilus by using API NH cards.The results of antimicrobial susceptibility showed that they were all sensitive to chloramphenicol,doxycycline,tetracycline,gentamicin,ofloxacin and ciprofloxacin.Conclusion The 10 isolates could be identified as Francisella philomiragia,so we should pay more attention to the infrequent pathogen for its inactive biochemical reaction and the misidentification by commercial detection systems.
6.Effect of idazoxan on permeability of blood-brain barrier and expression of MMP-9/TIMP-1 in mouse experimental autoimmune encephalomyelitis
Xinshi WANG ; Qingyi ZENG ; Zhenguo ZHU ; Pan ZHU ; Huiqin XU ; Rongyuan ZHENG
Chinese Journal of Pathophysiology 2014;(12):2254-2258
[ ABSTRACT] AIM:To study the effect of idazoxan ( IDA) on the permeability of blood-brain barrier ( BBB) and the expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in mouse ex-perimental autoimmune encephalomyelitis (EAE).METHODS: Female C57BL/6 mice (n=36) were randomly divided into control group, EAE group and IDA group, with 12 mice in each group.EAE was induced by myelin oligodendrocyte glycoprotein 35-55 ( MOG35-55 ) .IDA (2 mg/kg, ip, bid) was administered for 15 d after immunization.The neurological defects of the mice were observed daily and scored.The pathological changes were observed under microscope with HE stai-ning and LFB myelin staining.The BBB permeability was detected by Evans blue extravasation.The expression of MMP-9 and TIMP-1 in the brain of EAE mice was determined by Western blotting.RESULTS: Compared with EAE group, the score of neurological defects in IDA group was decreased, the inflammation was relieved, the BBB permeability was re-duced, and the expression MMP-9 and the ratio of MMP-9/TIMP-1 were decreased ( P<0.05 ) .CONCLUSION: The neuroprotective effect of IDA on mouse EAE might be related to the down-regulation of MMP-9 and the ratio of MMP-9/TIMP-1, thus reducing the degradation of BBB and the permeability of BBB, and ameliorating the pathologic process of EAE.
7.Effect of Yuhong ointment on the proliferation and differentiation of epidermal stem cells during rat wound healing
Huanyu KONG ; Fanfei KONG ; Qingyi HUANG ; Li LI ; Liping YANG ; Jia ZHU
Chinese Journal of Tissue Engineering Research 2007;11(46):9400-9403
BACKGROUND: Epidermal stem cells (ESCs) are of importance in the wound repair. Explaining the mechanism thatChinese herb speeds up the regeneration of injured skin from the angle of inducing stem cells deserves to be studied. OBJECTIVE: To observe the effect of Yuhong ointment on the proliferation and differentiation of ESCs during ratwound healing. DESIGN: A randomized controlled animal experiment. SETTING: Department of Pharmacology, Wangjing Hospital, China Academy of Chinese Medical Sciences. MATERIALS: This study was carried out in the Department of Pharmacology, Wangjing Hospital, China Academy ofChinese Medical Sciences between July and September 2006. Totally 114 healthy Wistar male adult rats, of cleangrade, weighing 180-210 g, were provided by the Laboratory Animal Center, Institute for Basic Theory of TraditionalChinese Medicine, China Academy of Chinese Medical Sciences were enrolled in this study. The processing ofanimals corresponded to the standard of Animal Ethics. Yuhong ointment was purchased from the PharmaceuticalCenter, Guanganmen Hospital, China Academy of Chinese Medical Sciences (Lot No. 20020110). Jingwanhong wasthe product of Tianjin Darentang Da'er Pharmcaceutical Co., Ltd (Lot No. Z12020440). METHODS: The 114 rats were completely randomly chosen and divided into 3 groups with 36 in each: Yuhongointment group, Jingwanhong group and model group, and the left 6 rats were involved as normal control group. Ratsin the normal control group were raised routinely, and no intervention was carried out. Immediately after beingmodeled, rats in the Yuhong ointment group were spread with Yuhong ointment, 0.1 g each wound, rats in theJingwanhong group were spread with Jingwanhong, 0.1 g each wound. Dressing change was daily carried out twice regularly until wound healing; The wounds of rats in the model group were untouched. MAIN OUTCOME MEASURES: On the 1st, 3rd, 5th, 7th, 14th and 21st days after modeling, wound healing time of rats in eachgroup was recorded, and integrinβ1 absorbance and transcription factor p63 positive cell amount were compared. RESULTS: Totally 114 rats were involved in the final analysis.① Wound healing time of rats in the Yuhong ointmentgroup and Jingwanhong group was significantly shorter than that in the model group, respectively (P < 0.05). ② Integrinβ1 absorbance and transcription factor p63 positive cell amount of modeled rats in the Yuhong ointmentgroup, Jingwanhong group and model group were significantly higher than those in the normal control group, respectively (P < 0.05); The above-mentioned two indexes in the Yuhong ointment group and Jingwanhong groupreached the peak on the 7th day, which was earlier than peak time in the model group, and peaked intensity of twoindexes was significantly higher than that in the model group, separately (P < 0.05). CONCLUSION: Yuhong ointment can promote rat wound healing, which may be associated with Yuhong ointmentinducing the proliferation and differentiation of ESCS left in the wound edge.
8.Rapid Diagnosis of Legionella pneumophial Pneumonia Used Polymerase Chain Reaction
Jun WANG ; Lianyou ZHENG ; Haitao PENG ; Zhaohui HU ; Yuanli LIU ; Qingyi ZHU
Chinese Journal of Nosocomiology 2006;0(08):-
OBJECTIVE To prove the diagnosis value for Legionnaires pneumophial pneumonia using polymerase chain reaction. METHODS L. pneumophial-DNA (LPN-DNA) from 47 spuum and 6 bronchoalveolar lavage fluid samples collected from 53 patients with atypical pneumonia was detected by PCR. RESULTS The positive rate of LPN-DNA in 53 patients with atypical pneumonia was 9.4%, while the positive rate of sputum and bronchoalveolar lavage fluid samples was 6.4%and 33.3%, respectively. CONCLUSIONS LPN-DNA detected by PCR for early diagnosis of atypical pneumonia has favorable clinical application.
9.Identification of a newly reported Francisella species by average nucleotide identity based on high-throughput whole genome sequencing technology
Lei ZHANG ; Minling ZHENG ; Ya WANG ; Haiyun CAI ; Guangyuan DENG ; Qingyi ZHU ; Cha CHEN ; Pinghua QU
Chinese Journal of Clinical Laboratory Science 2017;35(7):499-502
Objectives To identify the Francisella strain isolated from blood of a patient with drowning-associated pneumonia.Methods The whole genome of the strain,designated Wenzhou1,was sequenced using the high throughput sequencing technology by 2000/miSeq system of Illumina platform,and the obtained genome draft was assembled by MicrobeTrakr Plus software.The phylogenetic neighbors of Wenzhou1 were obtained by NCBI BLAST analysis from GenBank database for the gene sequences of 16S rRNA,malate dehydrogenase(mdh),DNA-directed RNA polymerase subunit beta (rpoB) and succinate dehydrogenase subunit alpha (sdhA).The average nucleotide identity(ANI) between Wenzhou1 and its phylogenetic neighbors was analyzed by the software OrthoANI using NCBI BLAST search under the Java Runtime Environment Version 8.Results The genome size of Wenzhou1 was 1.96 × 106 bp,containing 74 contigs.The genomic G + C mol% of Wenzhou1 was 32.1%,which was similar to the other species of genus Francisella and Allofranicella.Based on the analysis of NCBI BLAST of GenBank for the similarities of 16S rRNA gene,mdh gene,rpoB gene and sdbA gene sequences,Wenzhou1 was most closely related to F.hispaniensis FSC454 and Francisella cf.novicida 3523.The ANI of Wenzhou1 was 97.8% to F.hispaniensis FSC454,97.5% to 97.6% to Francisella cf.novicida 3523,but only 91.3% to 91.5% to the four subspecies of F.tularensis.Conclusion ANI analysis based on whole genome sequence should be an accurate,effective method for bacterial identification.Wenzhou1 could be identified as F.hispaniensis by ANI with high-throughput whole genome sequencing technology.
10.Identification and characterization of one Roseomonas strain
Lei ZHANG ; Pinghua QU ; Qingyi ZHU ; Huixia HU ; Shouyi CHEN ; Minling HU ; Zhaohui HU
Chinese Journal of Laboratory Medicine 2011;34(1):41-45
Objective To identify one runny mucoid-like Gram-negative bacteria with pink pigment isolated from clinical pus sample. Methods The pus sample was aseptically extracted from a deep lesions of one patient, then stored in Amies medium at room temperature for transportation. One sheep blood plate and one chocolate plate were used to detect the possible pathogens from the specimens. After inoculation, the plates were placed in a humidified incubator with 5% CO2 at 35 ℃. To identify the obtained isolates, we used the commercial Vitek2 and API systems, combining some traditional morphological examination and classical biochemical and physiological characteristics. For pure cultures, the cellular fatty acids were extracted, methylated, and determined by gas chromatography method. The 16S rRNA gene was amplified and sequenced by a commercial broad-spectrum PCR primers. The phylogenetic tree based on 16S rRNA gene was constructed by Mega 4.1 software using the neighbour-joining methods with 1 000 bootstrap replications. Results One runny mucoid-like Gram-negative bacterium, named K8756, was isolated both on sheep blood and chocolate plates after 72 h incubation. The API 20NE profile was 1245045 after a 3-day culture, which would be identified as Ochrobactrum anthropi with a good confidence of 98% probability. It was identified as Ralstonia pickettii and Bordetella bronchiseptica by VITEK 2 GN kits. However, further comparative 16S rRNA gene sequences showed that strain K8756 was closely related to the valid published Roseomonas mucosa MDA 5527 with 100% identity. Colonial morphologic features, phenotypic characteristics and major cellular fatty acid composition were also with high similarity to Roseomonas mucosa. Conclusions Strain K8756( = GIMCC 1.0030 ) is identified as Roseomonas mucosa by the polyphasic phenotypic and genotypic characteristics. The comparative analysis based on 16S rRNA gene sequences is a useful method for identifying the problematic and newly named bacteria.