Expression of microRNA-221/222 in patients with monoclonal gammopathy of undetermined significance and multiple myeloma / 浙江大学学报·医学版

To detect the expression of miR-221/222 in serum and plasma cells in patients with monoclonal gammopathy of undetermined significance(MGUS) and multiple myeloma(MM), and to explore the possibility of miR-221/222 as biomarkers in the diagnosis and prognosis predicting of MGUS and MM.Bone marrow and serum samples from 14 patients with newly diagnosed MGUS, 81 patients with newly diagnosed or relapsed MM and 10 controls were collected from Sir Run Run Shaw Hospital of Zhejiang University and Tongde Hospital of Zhejiang Province during January 2013 and December 2015. The expressions of miR-221/222 in serum and in sorted CD138 positive plasma cells were detected by qRT-PCR, and the relative expression of miR-221/222 (Δct) was compared between the groups. Serum levels of miR-221 before and after treatment were compared in both remission group (=22) and refractory group (=13) in MM patients, and its correlation with serum level of β-MG was assessed using Pearson's correlation analysis.Serum levels of miR-221/222 in MGUS and MM groups were significantly higher than those in control group (all<0.01), while miR-221/222 levels in plasma cells were significantly lower in MGUS and MM groups than those in the control group (<0.05 or<0.01). No significant difference in miR-221/222 levels in serum and plasma cells was observed between MGUS group and MM group (all>0.05). There was no correlation between miR-221/222 levels in serum and plasma cells (=0.024 and -0.127, all>0.05), but miR-221 levels were correlated with miR-222 levels in both serum and plasma cells (=0.534 and 0.552, all<0.01). Receiver operating characteristic (ROC) curves showed that the areas under the curve (AUCs) of serum miR-221/222, plasma cell miR-221/222 in diagnosis of MGUS/MM were 0.968, 0.976, 0.801 and 0.727, respectively. There was no significant difference in serum level of miR-221 among MM patients with different paraprotein isotypes (>0.05), but serum level of miR-221 in patients with relapsed MM was higher than that in patients with newly diagnosed MM (<0.01). Compared with the patients with MGUS or MM stageⅠ and Ⅱ, patients with MM stage Ⅲ were of higher serum levels of miR-221 (<0.01). Serum level of miR-221 decreased after chemotherapy in the remission group (=51.5,<0.01), but such decrease was not observed in the refractory group (=67.5,>0.05). Serum level of β-MG was positively correlated with serum level of miR-221 (=0.524,<0.01).miR-221/222 in serum and plasma cells may be biomarkers for early diagnosis of MGUS, and are helpful for diagnosis and efficacy evaluation of MM.

Evaluation of VCS technology in diagnosis for acute leukemia and its subtypes / 中华检验医学杂志

Objective To evaluate clinical application of VCS technology in diagnosis for acute leukemia and its subtypes by LH750 automated hematology analyzer. Methods A total of 178 leukemia patients (105 acute granilocytic leukemia, 42 acute lymphocytic leukemia, 31 acute monecytic leukemia)and 151 non-leukemia patients (86 granulocytic abnormal cases, 35 lymphocytic abnormal cases,30 monocytic abnormal cases) were enrolled into this study. Peripheral blood samples were analyzed by both Coulter LH750 automated hematology analyzer and manual microscopic examination. Microscopic examination was used as reference to evaluate the diagnosis value of VCS technology in blast detecting. VCS parameters were compared for AGL subsets. The clinical value of VCS technology in diagnosis for different acute leukemia was also evaluated by combining VCS scattergram and suspect flags provided by LH750. Results Compared with manual examination, a sensitivity of 95.5% (170/178) and specificity of 95.4% (144/151) were achieved by VCS technology in detecting blasts in acute leukemia and nonleukemia patients. In patients with AGL, the MNV, MNV-SD, MNC and MNC-SD (224.40 ± 23.37,37.40 ±12. 31,145. 80 ±7. 93,24. 79 ±5. 18) were higher than granulocytic abnormal patients( 169.96 ±11.50,29.21 ± 5.27,133.30 ± 5.50, 10.62 ± 4.09) and the differences were statistically significant ( t values were 16. 832, 5. 148, 5. 735, 19. 953 respectively, P < 0. 01 ). On the contrary, MNS ( 122. 90 ±6. 35) in AGL patients was significantly lower than granulocytic abnormal patients( 131.00 ± 5. 69, t =-7.64, P<0.01). In patients with ALL, the MLV, MLV-SD, MLC, MLS, MLS-SD (97.60 ± 13.40,22. 35 ± 7.94,110. 00 ± 4. 60,77. 60 ± 19.00,20. 61 ± 3.30 ) were higher than lymphocytic abnormal patients(82. 10 ± 3.00, 14.41 ±2.35, 100.60 ±2.70, 48. 10 ±3.50, 17.60 ± 1.60). The data also showed a statistical significance (t values were 7.576, 6. 118, 4. 041, 9. 353, 2. 988 respectively, P <0.05). In patients with AMOL, the MMV, MMV-SD, MMC, MMC-SD, MMS(197. 30 ±20.50,30.47 ±6.58,123.20 ± 10. 10,6.57 ± 1.57,98.00 ± 5.60) were significantly higher than monocytic abnormal patients( 167.80 ± 15.77,21.90 ± 9. 64,113.60 ± 6. 73,5.20 ± 3. 21,84. 20 ± 14. 35 ) and the differences were statistically significant (t values were 6. 332, 4. 033, 4. 650, 2. 993, 6. 273 respectively , P <0. 01 ).The differences of MNV, MNV-SD, MNC, MNC-SD, MNS-SD for different AGL subtypes were statistically significant ( F values were 21.2, 169. 5, 13.6, 3. 6 and 98.6 respectively, P <0. 01 ). For AGL, ALL and AMOL, the ratios of normal blast distribution in scattergram were 81% (85/105), 74% (31/42) ,and 74% (23/31). Conclusions The VCS parameters can reflect the changes of peripheral white blood cell morphology sensitively. VCS technology can diagnose acute leukemia and its subtypes efficieutly.

Research on detection of Legionella pneumophila by real-time fluorescence PCR / 中华微生物学和免疫学杂志

Objective To develop a real-time fluorescence quantitative PCR method for detection of Legionella pneumophila as a tool for environmental and clinical examination. Methods A pair of degen-erated primers and one TaqMan-MGB probe were designed to test the conserved region at the macrophage in-fective potentiation (mip) gene of Legionella pneumophila. TaqMan MGB real-time quantitative PCR assay was developed with pMD-19T plasmid including mip gene of Legionella pneumophila as standard sample. The sensitivity and specificity of the real-time quantitative PCR was evaluated using the standard sample and dif-ferent strains. Results The detection limit of 0.71 copy/μl was obtained for the standard sample in a reac-tion system of 0.6μl of sense and antisense primers (20μmol/L), respectively, 0.4μl of probe (20μmol/L) and 6.0μl of DNA temple. Conditions for the PCR reactions were as follows. After an initial de-naturation at 95℃ for 20s, 40 amplification cycles were performed. Each cycle consisted of denaturation at 95℃ for 10 s, primer annealing at 50℃ for45s. The PCR Ct value of a standard strain and 12 isolates was in a scale of 13.23 and 16.04. However, the Ct values of the strains of Staphylococcus aureus, Salmonella typhimurium, Vibrio parahaemolyticus, Escherichia coli, Pseudomonas aeruginosa and Shigella sonnei were greater than 30. Conclusion The real-time quantitative PCR method has good sensitivity and specificity and the result has potentiality of applying for detecting Legionella pneumophila.