1.Isolation, identification and drug sensitivity analysis of Mycobacteroides abscessus in a hospital in Hainan Province from 2014 to 2021
WANG Jieying ; CHEN Zhuolin ; XU Yuni ; YU Chunchun ; KONG Fanrong ; CHEN Qiong ; ZHONG Yeteng
China Tropical Medicine 2023;23(9):947-
Abstract: Objective To identify the species of Mycobacteroides abscessus complex (MABC) in patients with pulmonary infection from the Second Affiliated Hospital of Hainan Medical University, and to investigate the species types, drug sensitivity and population distribution of MABC in pulmonary infection in Hainan. Methods Respiratory tract specimens were collected from suspected tuberculosis patients who visited the Second Affiliated Hospital of Hainan Medical University from January 2014 to December 2021 and cultured for Mycobacterium isolation. Non-tuberculous mycobacteria (NTM) strains were preliminarily identified by p-nitrobenzoic acid/thiophen-2-carbohydrazide (PNB/TCH) medium and DNA microarray chip, and then MABC and its subspecies were identified by hsp65 and rpoB gene sequencing. In vitro antimicrobial susceptibility test was performed by broth microdilution method. Results A total of 3 025 respiratory specimens from suspected pulmonary tuberculosis patients were collected during the study period. Among the 123 patients with identified MABC isolates, 124 MABC strains were isolated and identified, including 74 strains of Mycobacteroides abscessus subsp. abscessus, 38 strains of Mycobacteroides abscessus subsp. massiliense and 12 strains of Mycobacteroides abscessus subsp. bolletii. Among them, 118 patients had single MABC subspecies infection, one patient had mixed infection with two MABC subspecies, two patients had mixed infection with MABC and other NTM, and two cases had mixed infection with MABC and M.tuberculosis. There were more female patients than male patients with a ratio of 1:0.64, and those aged 50 and above amounted to 76.42% (94/123, 95%CI: 67.93%-83.61%). There was no significant difference in age distribution between male and female patients (Z=-0.944, P=0.347). The drug susceptibility results showed that all MABC strains were sensitive to Tigecycline (TGC), with a resistance rate of 0.81% (1/124) to Amikacin (AK), and resistance rates of 6.45% (8/124), 32.26% (40/124), and 74.19% (92/124) to Cefoxitin (FOX), Linezolid (LZD), and Imipenem (IPM), respectively. For Clarithromycin (CLR), MABC showed induced resistance , and there was a statistically significant difference in the CLR (14D) resistance rates among the three subspecies (χ2=66.335, P<0.001). The resistance rates to Tobramycin (TOB), Doxycycline (DOX), Moxifloxacin (MFX), Ciprofoxacin (CIP), Trimethoprim/Sulfamethoxazole (TMP-SMX), and Amoxicillin/Clavulanic acid (AMC) were high, all >80%. Conclusion In Hainan Province, pulmonary infections with MABC are mainly caused by Mycobacteroides abscessus subsp. Abscessus, which show high rates of inducible resistance to CLR. Timely and accurate identification of MABC to subspecies and drug susceptibility testing are of significant important for clinical decision-making.
2.The combination of flow cytometry and automated cell image analyzer in determining the nature of ascites and pleural effusion nature
Yuni GUO ; Shumin XU ; Jinyan DUAN ; Wenyan CHEN ; Ruibing LI ; Zhengguan WANG ; Fan ZHANG ; Ce WANG ; Chengbin WANG
Chinese Journal of Laboratory Medicine 2015;(3):183-185
Objective To explore the clinical value of flow cytometry( FCM) and DNA automated cell image analyzer ( AICM) in determine the character of ascites and pleural effusion.Methods This was a cross-sectional study.203 ascites and pleural effusionsamples were random selected from PLA hospital inpatients between August 2013 to June 2014 .The DNA content of sediment cells were detectedthrough the FCM and AICM respectively benign and malignant disease were differentiated according the counts and proportion of aneuploid cells.The sensitivity, specificitywere calculated byROC curves.Results The sensitivity, specificity and accuracy of flow cytometry cell in detectingtumor cells were 78.6%,80.0% and 79.2%%, while the sensitivity, specificity and accuracy of image analyzer were 83.5%,78.6% and 81. 3%respectively.When FCM and AICMwere combined ,the sensitivity, specificity and accuracyincreased to 92.2%, 86.3% and 89.6%.Conclusions Compared toconventional cytology test, the sensitivity and specificity were significantly high when the two methods were combined .Therefore, the combination method can be used to assist in clinical identification of the nature of ascites and pleural effusion and to help the diagnosis of disease.
3.Molecular mechanism of tetramethylpyrazine to induce human promyelocytic HL-60 leukemia cells differentiation.
Yuni WU ; Youhua XU ; Xiaoyan GU ; Yanni HU ; Cuicui WANG
China Journal of Chinese Materia Medica 2011;36(21):3007-3011
OBJECTIVETo study the molecular mechanism of tetramethylpyrazine to induce human promyelocytic HL-60 leukemia cells differentiation.
METHODThe cell proliferation was determined by MTT. The differentiation of the cells was detected by NBT reduction test. Cellular morphology was observed by Wright's staining. Cell cycle distribution and the distribution of CD11b, CD14 were detected by flow cytometry. Then RT-PCR and Western blot assay were employed to detect the expressions of c-myc, p27, CDK2 and cyclinE1 in HL-60 cells after exposure to TMP.
RESULTTMP inhibited the proliferation in a dose and time dependent manner. TMP at the concentration of 200 mg x L(-1) to 300 mg x L(-1) induced unterminal differentiation of HL-60 cell and synergistically blocked the cell cycle progression of HL-60 cells in G0/G1 phase. The expression of c-myc was down-regulated as well as the protein expression of cyclin E and CDK2, while the mRNA and protein expression of P27 were remarkably up-regulated.
CONCLUSIONSmall doses of TMP induces differentiation of HL-60 cells throughout the cell cyde, as detected by a slower rate of accumulation in G0/G1, possibly by regulating the expression and activity of G1/S phase-related molecules.
Cell Cycle Proteins ; genetics ; metabolism ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Leukemic ; drug effects ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; genetics ; metabolism ; physiopathology ; Pyrazines ; pharmacology
4.SE-iFISH used in the detection of circulating tumor cells in patients of hepatocellular carcinoma
Jun SHI ; Guangqiang YE ; Yuni KE ; Chunhui YE ; Shiwei YANG ; Xuan TONG ; Jing XU ; Jiahong DONG
Chinese Journal of General Surgery 2018;33(10):865-868
Objective To investigate the correlation between patients clinical characteristics and the number and subtype of circulating tumor cells (CTCs) from peripheral blood of perioperative hepatocellular carcinoma (HCC)patients by SE-iFISH.Methods 20 HCC patients undergoing radical resection were enrolled from June 2015 to June 2016.The SE-iFISH technique was used to separate and identify circulating tumor cells.The pathology and clinical data were used to evaluate patients survival in combination with CTCs characteristics.Results A total of 347 CTCs were detected,of which 114 were triploid,64 were tetraploid,and 165 were pentaploid.The number of preoperative CTCs and the number of preoperative triploids was significantly correlated with the presence of vascular tumor emboli (Z1 =-2.080,P =0.037,Z3 =-2.321,P =0.020) and TNM staging(Z2 =-2.148,P =0.032,Z4 =-2.526,P =0.012).Postoperative patients disease-free survival in high CTCs detection group was significantly shorter than that of CTCs low expression group (x2 1 =7.486,P =0.006,x22 =12.056,P =0.001).Conclusion Detection of the number and the specific subtypes of CTCs with SE-iFISH strategy in patients with HCC help predict treatment efficacy and prognosis.