1.Clinical features and prognosis analysis of gallbladder cancer after radical resection in elderly patients
Zhijian WEN ; Junli HUANG ; Xiaogang XIA ; Bin LIU ; Pinghua YANG ; Baohua ZHANG
International Journal of Surgery 2018;45(10):673-679,封3
Objective To clarify the characteristics and postoperative benefit of gallbladder carcinoma in elderly patients (≥ 65 years old).Methods Two hundred and seventy-three patients of gallbladder carcinoma were collected,who were treated intent resection from January 2004 to December 2012 in the Department of Hepatobiliary Surgery,Eastern Hepatobiliary Hospital,Second Military Medical University.More than 65 years old was defined as the elderly,else was defined as the younger.The clinical-pathological features and prognosis of 85 elderly patients(elderly group) and 188 younger patients (younger group) were retrospectively analyzed.The survival of patients were followed up by telephone or outpatient.The incidence of hypertension,incidence of diabetes,TNM staging,and median CA19-9 were compared between the two groups.Continuous variables using a two-sample t test or Mann-Whitney U test.Categorical variables were compared by the Chi-square test or Fisher probability method.The survival curve was drawn by the Kaplan-Meier method.The univariate analysis and multivariate analysis of prognosis were respectively done using the Log-rank test and COX regression model.Results The incidence of hypertension,incidence of diabetes,TNM stage ratio (Ⅲ + Ⅳ/Ⅰ + Ⅱ),and CA19-9 median in the elderly group were 30.6%,11.8%,27.6 and 69.3 U/ml,respectively.The differences in the younger group were 13.8%,4.8%,7.9 and 28.2 U/ml,respectively,with statistically significant difference between the two groups (all P < 0.05).The incidence of complications was 54.1% in the elderly group and 48.9% in the younger group,with no significant difference between the two groups (P =0.302).The median survival of the elderly group was 28.01 months,and the median survival of the younger group was 36.20 months,with no statistical difference between the two groups (P =0.131).Cox analysis showed that independent prognostic risk factors for the elderly patients with gallbladder cancer included liver invasion (HR =2.386,95% CI:1.379-4.127,P =0.002) and lymph node metastasis (HR =1.866,95 % CI:1.100-3.167,P =0.021).Conclusions Radical resection is safe and feasible for elderly patients with gallbladder carcinoma.Age is not a contraindication for surgery.Radical resection can get the same benefits as young people.Liver invasion and lymph node metastasis are independent risk factors affecting the prognosis of the elderly patients with gallbladder carcinoma.
2.Identification and classification of non-tuberculosis Mycobacterium by matrix-assisted laser desorption ionization-time of flight mass spectrometry
Song LI ; Yanjiao YAO ; Pinghua QU ; Cha CHEN ; Weizheng ZHANG ; Qiwei LI ; Zhenjie XU ; Fu CHEN
Chinese Journal of Laboratory Medicine 2018;41(2):109-115
Objective Reference standard of the RPOB(rifampin resistance)gene recommended by CLSI-MM18A(Interpretive Criteria for Identification of Bacteria and Fungi by DNA Target Sequencing) was used to evaluate the ability of MALDI-TOFMS techniques for the identification and classification of non-tuberculous Mycobacterium.Methods Fifty five clinicalstrains were collected from 2012 to 2016 with different sources.The RPOB gene was sequenced, and results were applied to phylogenetics analysis. MALDI-TOF MS technology was implemented to identify the strains, and cluster analysis was conducted based on protein fingerprint.The consistency of two methods for NTM identification and typing was evaluated.Results The RPOB gene method showed a good ability of identification(similarity>99.0%) and subtyping(to subspeciesof the complex level).The French BioMérieux MALDI-TOF MS identified 89.1% of 55 strains to genus level and 78.2% to species level.The phylogeneticsanalysis of protein fingerprint by SARAMS Premium software also showed good typing ability.Conclusions MALDI-TOF MS technology can identify and classify non-tuberculous Mycobacterium effectively,which is rapid and easy.It is complementary to RPOB gene method in laboratory application.
3.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.
4.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.
5.Correlation between ponticulus of atlas and abnormal morphology of V3 segment of vertebral artery using multi-slice spiral CT angiography
Pinghua WAN ; Benxing WANG ; Yusheng LIAO ; Daiping GAO ; Fukang ZHANG
Chinese Journal of Radiology 2016;50(9):657-661
Objective To explore the correlation between ponticulus of atlas and abnormal morphology of V3 segment of vertebral artery. Methods All patients included in this study were underwent CTA and 3D reconstruction examination of intracranial ponticulus and cervical in our workstation, as well as the site, shape of petals and its relationship with vertebral artery was observed. The diameter, shape and trail of vertebral V3 segment were measured and analyzed. The results were analyzed byχ2 test. Results A total of 195 sides (157 cases) ponticulus were detected in 613 cases (1 226 sides), the abnormal morphology of V3 segment of vertebral artery were 103 sides and combined with ponticulus 69 sides among them, including (according to side): the collectivity was 8.4% (103/1 226), combination ponticulus was 35.4% (69/195), uncombination ponticulus was 3.3%(34/1 031), and the risk with ponticulus was increased compared with no ponticulus, and showed statistical significance by Chi-square test(χ2=215.679,P=0.001). The difference of abnormality of vertebral artery was performanced in number, site and shape of ponticulus, we found that the ponticulus composite was 57.1%(12/21) and higher than ponticulus simple 32.8%(57/174)(χ2=4.873,P=0.027), the ponticulus borderl was 68.8%(11/16) which was also higher than ponticulus posticus(27.2%) and ponticulus lateralis(36.4%)in the ponticulus simple(χ2=11.357,P=0.030), the sharp ones (64.2%) were higher than the blunt(14.6%)in the tip of bony processes of the incomplete type ponticulus(χ2=26.813, P=0.001),the smaller ones were higher than the larger in the distance of the gap between the two tips of bony processes in the incomplete type ponticulus(χ2=9.212,P=0.010), and the smallerones(87.5%)were also higher than the larger(28.6%)in the diameter of the hole of complete type ponticulus(χ2=18.193,P=0.001), all of the above ones showed statistical significance among all groups by Chi-square test (P<0.05). Conclusions Ponticulus of atlas can significantly increase the risk of abnormal morphology of V3 segment of vertebral artery, and the differences of abnormality of vertebral artery were performed in number, site and shape of ponticulus, which included the ponticulus composite, ponticulus borderland, the shape tip of bony processes, the moderate distance between the two lip of bony processes, and the smaller diameter of complete type Ponticulus.
6.Biotype identification and epidemiological analysis of twenty-four Brucella strains
Lei ZHANG ; Pinghua QU ; Shangwei WU ; Jingdiao CHEN
Chinese Journal of Endemiology 2016;35(4):251-255
Objective To identify the biotype and analyze the epidemiological characteristics of twentyfour Brurella strains from the primary hospitals in Guangdong Province.Methods The twenty-four Brucella strains,collected from Oct.2009 to Oct.2015,were identified by routine biochemical methods,VITEK 2 COMPACT automatic microbial identification analyzer,16S rRNA gene sequencing and biology phenotype based on serological and bacteriophages lysis test.The etiology was analyzed based on clinical data,biotypes of the isolates and other clinical information.Results All of the twenty-four strains were Gram-negative coccobacilli,including two strains of Brucella suis biotype Ⅱ,four strains of Brucella melitensis biotype Ⅰ and eighteen strains of Brucella melitensis biotype Ⅲ.By GN card of VITEK 2 COMPACT automatic microbial identification analyzer,one strain was mistaken as Bordetella bronchiseptica and two strains were mistaken as Ochrobaetrum anthropi.The 16S rRNA gene sequencing showed they were high homology to Ochrobactrum intermedium and Ochrobaetrum anthropi,which completely excluded the possibility of Bordetella bronchiseptica.Tbe clinical data showed that all of the twenty-four patients were adults with an average age of 49.0 years old,men and women were twelve people respectively,with no significant gender differences and no occupational exposure,which presenting a wide and diverse range of nonspecific clinical signs and symptoms,but brucellosis was not aware of by the physician.Conclusion Brucella melitensis biotype Ⅲ is the main pathogen of brucellosis,with the characteristics of sporadic outbreak and occult infection in the primary hospitals in Guangdong Province.
7.Determination of Plasticizer DEHP Released from Medical Injection Equipment.
Jian ZHENG ; Minzhu WANG ; Wang ANYAN ; Pinghua XU ; Yin HAN ; Yan WEN ; Li ZHANG
Chinese Journal of Medical Instrumentation 2015;39(6):451-453
To established an effective GC-MS /MS method for the contents determination of the residual DEHP in injection equipment, and investigate the effect of the pretreatment on the measurement. To simulate the clinical conditions of use, under the condition of 37 degrees C balance extraction, extract liquor by chloroform extraction, then the extract followed by analysis of GC-MS /MS. The method was simple, rapid, sensitive and accurate. The limits of quantitation (LOQ, S/N = 5) of cyclohexanone was 0.075 μg/mL, The spiked average recoveries ranged from 92% to 98%. The relative standard deviations (RSDs) of the method ranged from 1.01% to 1.61%, The method was simple, fast, sensitive and accurate, and may serve as a mass control method for residual DEHP in injection equipment.
Cyclohexanones
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chemistry
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Diethylhexyl Phthalate
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chemistry
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Equipment Contamination
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Gas Chromatography-Mass Spectrometry
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Injections
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instrumentation
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Limit of Detection
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Plasticizers
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chemistry
9.Study of anti-fatigue effect of methylphenidate hydrochloride oral fast dissolving films and its mechanism
Lihong ZHANG ; Pinghua XU ; Na WU ; Chengying SHEN ; Hailong YUAN ; Jin HAN
Journal of Pharmaceutical Practice 2015;(6):522-524
Objective To study the anti-fatigue effect of methylphenidate hydrochloride oral fast dissolving films (MPH-OFDF) and its mechanism .Methods 60 mice were randomly divided into 6 groups as:normal control group (physiological sa-line) ,model group (physiological saline) ,Yiqiyangxue oral liquids positive group (7 .00 mg/kg) ,MPH-OFDF high-dose group (5 .20 mg/kg) ,MPH-OFDF middle-dose group (2 .60 mg/kg) and MPH-OFDF low-dose group (1 .30 mg/kg) .Besides the normal control group ,model group and positive group were orally administered ,the other groups are administered with the drug once daily sublingually daily for consecutive 15 days .The mice were put in the load-weighted swimming test 30 min after the last oral administration ,then the anti-fatigue effect was assessed based on recording exhausting swimming time and detec-ting the levels of serum lactale dehydrogenase (LDH) ,creatine kinase (CK) ,triglycerides (TG) in mice .Results Compared with control group ,the middle-dose and the high-dose MPH could prolong the exhausting swimming time (P<0 .05 ,P<0 .01) and decrease the activity of LDH and CK significantly (P<0 .05 ,P<0 .01);in addition the middle-dose MPH could decrease the content of TG (P<0 .05) .Conclusion The MPH had marked anti-fatigue effect that may be associated with reduced ser-um LDH ,CK and TG .
10.Identification of Streptobacillus moniliformis isolated from the knee joint pus by 16S rRNA gene sequencing and biochemical reactions
Weizheng ZHANG ; Guangyuan DENG ; Pinghua QU ; Wenke CHEN ; Dongling LIN ; Cha CHEN
The Journal of Practical Medicine 2014;(11):1814-1817
Objective To indentify Streptobacillus moniliformis isolated from the knee joint pus by 16S rRNA gene sequencing and biochemical reactions and explore the clinical value of the method. Methods The bacterial 16S rRNA gene sequence-based identification, bacterial morphology, VITEK 2 automate systems, API 20NE strips, API 20E strips and API 50CH were performed to identify the rare bacteria. Results The bacteria grew slow on blood agar and chocolate agar and were inhibited on Maconkey agar. The bacterial colony on blood agar tookes the form of 1~2 mmomelette, which was translucent and moist with circular protrusion and smooth edges. They were Gram-staining negative and in catenation, its thalli 1~3μm, round, oval or fusiform. Vitek 2 GN-13, API 20NE and API 20E were unable to reach the identification of the bacteria. 16S rRNA gene sequencing showed the bacteria were similar to streptobacillus moniliformis by 100%. Conclusion The rare bacteria isolated from left knee joint are streptobacillus moniliformis. 16S rRNA gene sequences combined with the biochemical reactions is accurate in the identification of these bacteria.

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