Humans ; Multiple Myeloma/genetics* ; Karyotyping ; Translocation, Genetic/genetics*

Objective:To evaluate the potential differential diagnostic value of QuantiFERON-TB Gold Plus(QFT-Plus) in patients with active tuberculosis (ATB) and latent tuberculosis infection (LTBI) people.Methods:Case-control study. A total of 108 healthcare workers and 30 ATB patients in Xi′an Chest Hospital were tested by QFT-Plus from April to November 2019, and the demographic characteristics were analyzed.Then, flow cytometry was used to analyze the relations between the QFT-Plus TB2-TB1 and the distribution of peripheral blood T lymphocyte subsets in ATB patients with positive culture results.Finally, with 34 QFT-Plus positive volunteers as LTBI group and 30 bacteriologically confirmed ATB patients as ATB group, the QFT-Plus new lyadded antigen and its potential differential diagnostic value between LTBI and ATB groups was evaluated by using the receiver operating curve (ROC).Results:In patients with ATB,QFT-plus TB2-TB1 was positively correlated with the proportion of CD8+T cells in peripheral blood T lymphocytes( r=0.586, P=0.004), negatively correlated with the proportion of CD4+ T cells( r=-0.511, P=0.015) and the ratio of CD4/CD8 ( r=-0.520, P=0.013).The peripheral blood TB2-TB1 in the ATB patients was significantly higher than that in the LTBI group[0.47(0.12,1.17) IU/ml versus 0.01(-0.08,0.22) IU/ml, U=233.5, P<0.001]. QFT-Plus TB2-TB1 can effectively distinguish ATB from LTBI, with an area under the ROC curve of 0.771 (95 %CI=0.653-0.889, P<0.001). Conclusion:QFT-Plus specific CD8 response (TB2-TB1) has the potential value to identify ATB from LTBI people.

Objective: To investigate the effects of prostate cancer exosomes on the migration and invasion ability of stromal cells (WPMY-1), and to explore its mechanism. Methods: Exosomes in LNCaP-AI+F prostate cancer cell supernatant were isolated by ultracentrifugation and the typical structure of exosome was captured by electron microscope. The particle size distribution was analyzed by Zetaview, and Wb was used to identify the marker proteins and other proteins.After co-incubation of WPMY-1 cells and prostate cancer exosomes (40 µg/ml), laser confocal microscope was used to observe the uptake of PKH67-labeled exosomes by WPMY-1 cells; Transwell assay was used to detect the migration and invasion ability of WPMY-1 cells; qPCR was performed to detect the expression of three cancer-associated fibroblast (CAF)-related molecules (IL-8, PDGFB and MMP9) at mRNA level; and the phosphorylation of EGFR and ERK1/2 was analyzed by Wb. Results: Typical cup-shaped structure of exosomes was observed under electron microscope. The Zetaview results showed that the particle size distribution was concentrated at about 100 nm. The expression of exosome marker proteins CD63 andALIX further verified that the isolated particles were exosomes. Besides, EGFR, HER2 and SRC, which were related to the progression of prostate cancer, were also enriched in exosomes. After co-incubation, confocal microscope imaging showed a number of PKH67 labeled exosomes in recipient WPMY-1 cells. Transwell experiments showed that exosomes could significantly enhance the migration and invasion ability of WPMY-1 cells (all P<0.01). Compared with the control group, increased secretion of IL-8, PDGFB and MMP9 was observed after exosome treatment (40 µg/ml) (P<0.05 or P<0.01). Wb indicated that exosomes could promote the phosphorylation of EGFR and ERK1/2 of WPMY-1 cells (P<0.01). Conclusion: Prostate cancer cell exosomes could act on the stromal cell WPMY-1 to highly express multiple CAF-related molecules, promote the phosphorylation of EGFR and ERK1/2 and enhance the migration and invasion ability of WPMY-1 cells.

Objective: To study the effects of Xiaoyaosan on sexual behavior and inflammatory factors of rat with depression. Methods: The establishing depressive rat models of chronic mild unpredictable stress were established. The rats were randomly divided into model group and Xiaoyaosan group, 15 in each group. The Xiaoyaosan group was given 4.612 5 g/(kg•d) Xiaoyaosan, the blank group and the model group were intragastrically administered with normal saline once daily. After 4 weeks of oral administration, mount frequency and perineal licking numbers to female rats in each experimental group were observed by sexual behavior experiment, and the spleen index and thymus index were calculated. The serum DA, IL-1β, IL-6 levels were tested by Elisa. Results: Compared with the normal group, the mount frequency (4.42 ± 3.91 vs. 11.58 ± 6.54), perineal licking numbers (3.53 ± 3.29 vs. 16.36 ± 10.68) and serum DA levels (14.14 ± 0.71 pg/ml vs. 17.44 ± 4.06 pg/ml) in the model group significantly decreased (P<0.05 or P<0.01), and the thymus index (0.012 4 ± 0.001 8 vs. 0.009 3 ± 0.001 7), serum IL-1β (39.45 ± 11.98 pg/ml vs. 28.62 ± 6.61 pg/ml), IL-6 levels (9.74 ± 1.49 pg/ml vs. 6.40 ± 0.66 pg/ml) significantly increased (P<0.05 or P<0.01). Compared with the model group, the contents of serum DA in Xiaoyaosan group (15.90 ± 1.24 pg/ml vs. 14.14 ± 0.71 pg/ml) increased significantly (P<0.01), and the thymus index (0.009 9 ± 0.001 3 vs. 0.012 4 ± 0.001 8) and IL-6 levels (8.32 ± 0.39 pg/ml vs. 9.74 ± 1.49 pg/ml) decreased significantly (P<0.05). Conclusions The Xiaoyaosan had a therapeutic effect on immune dysfunction and increases the expression of serum dopamine in depressive rats, which may be one of the mechanisms of its anti-depression.

Objective: To optimize the existing methods of isolation and purification for exosomes from urine and explore the effects of different storage conditions on the content of exosomal RNA in urine. Methods: The exosomes in human urine samples were extracted by different precipitation method, i.e., precipitation following first concentrating and direct precipitation, respectively, and the separation efficiency and cost of the two methods were compared. ExoQuick-TCTM precipitation kit was used to extract exosomes. Nanoparticle tracking analysis technique (NTA) was used to detect the concentration and particle size distribution of exosome. Dynamic light scattering (DLS) was used to detect the potential of exosome. Transmission electron microscopy (TEM) was used to observe morphology of exosomes. western blot was used to analyze the exosomal marker molecules CD63 and Alix. The extraction method of the precipitation following first concentrating was used to verify the reliability of the optimized method in 10 clinical urine samples . Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of exosomal RNA marker let-7c and PSA mRNA in the urinary exosomes from 20 patients with prostate cancer after repeated freeze-thaw (0 [i.e., fresh], 1 , 3 and 5 times) and 9 patients with prostate cancer frozen at -80 ℃ for different time (0 [i.e., fresh], 1, 2 and 4 weeks), and were statistically analyzed by Wilcoxon rank sum test for differences between the 2 groups. Results: The size distribution of exosomes extracted by the two methods was 30 to 150 nm by NTA, both of which were displayed as single peaks. The results of DLS showed that the potentials of exosome extracted by the two methods were negative values. The size of the exosomes extracted by the two methods was consistent observed under TEM namely the diameter distribution was 30 to 150 nm. western blot analysis confirmed that CD63 and Alix, the exosome labeling molecules, existed in the optimized method. The concentration of exosomes extracted from the 10 urine samples all reached 10 9 to 10 11 particles/mL. The contents of let-7c and PSA mRNA in exosomes decreased significantly after 5 freeze-thaw cycles, and the Z values were -1.79 and -1.73, respectively (P＜0.05). The RNA content of the exosomes remained stable after freezing at -80 ℃ for 1 month. Conclusion The optimized exosome extraction method could reduce greatly the cost under the premises of ensuring the concentration and quality of exosomes. The isolated exosomes may keep stable RNA content after freezing at -80 ℃ for a short time, but could not be frozen and thawed repeatedly for more than 5 times.

Objective Studying the differential expression of exosomal miRNAs between androgen-dependent prostate cancer cell and androgen-independent prostate cancer cell,in order to further elucidate the mechanism of androgen-independent prostate cancer and find new targets for its treatment.Methods Prostate cancer androgen-dependent LNCaP cell and prostate cancer androgen-independent LNCaP-AI + F cell (induced by androgen and flutamide) were selected as the study subjects.Illumina HiSeqTM 2500 was used to perform high-throughput sequencing between the two groups.The differentially expressed exosomal miRNAs was verified by quantitative real-time PCR(qRT-PCR).The difference was statistically significant by t-test.Results Through the analysis of high-throughput sequencing results,thirteen molecules were screened increased in extracellular exosomes of the androgen-independent cell,including miR-7-5p,let-7a-5p,miR-375,miR-423-3p,miR-378a-3p,and miR-92a-3p,etc.Among them,miR-7-5p was verified by quantitative real-time PCR to be up-regulated by 19.52-fold (t=9.857,P=0.001).Conclusion Differentially expressed exosomal miRNAs may predict the development of androgen-independent prostate cancer and may further regulate the development of androgen-independent prostate cancer.

This review, based on the origin and the mechanism of release of exosomes, described the presence of HBV and HCV biomarkers in the exosomes and the mechanisms of secretion, and discussed the viral transmission, vaccination protection, and the influences on treatment and detection.Through basic research data, it preliminarily demonstrated the importance of nucleic acid detection being a gold standard, and pointed out that another new cause of the occult infection was existence of HBV and HCV in the exosomes.(Chin J Lab Med, 2018, 41: 496-498)

Objective: To discuss the application of matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) in the identification of Aspergillus and evaluate its performance. Methods: the clinical isolates of Aspergillus collected from May 2017 to March 2018 in PLA General Hospital were identified by VITEK MS V3.0 and the results were analyzed. The ITS sequencing resultswere used as the gold standard. Results: It identified 9 Aspergillus species (including 12 Aspergillus species in total) through the V3.0 database, accounting for 86.24% of the total clinical isolates. The identification rate by VITEK MS was 91.49% with 16.51% was not identified. The coincidence rate of genus was 93.62%, of which only two Aspergillus versicolor were identified to the level of the genus. According to the confidence level analysis, 88.30% of the strains obtained more than 99% of the identification rate. 13.83% of the strains did not have the identification results for the first time, with the error rate of 3.19%. After secondary extractions, the percentage of unidentified strain was reduced to 6.38%, and the identification error rate was reduced to 2.13%. Combined with traditional identification and VITEK MS identification, the correct rate of strains identification was 98.94% on genus level, and was 93.62% on species level. The influence of other fungi on Aspergillus identification was 0%. Conclusion As a powerful supplement to the traditional identification method, MALDI-TOF MS showed a lot of convenience when applied in the identification of Aspergillus, which improves the identification accuracy and the identification ability for fungi in laboratory.(Chin J Lab Med, 2018, 41: 577-582)

Objective To shorten the turn around time of positive blood culture results by optimizing the blood culture positive specimen processing flow.Methods In January 26,2015,the microbiology department started the blood culture positive specimen processing flow optimization project,and applied the Lean Six Sigma method in the microbiological process management.The TAT data of 124 positive blood cultures containing Enterobacteriaceae were collected before and after the start of the project in about two months.We analyzed the turnaround time median,mean and standard deviation and reference Z value,process performance index,millions of error opportunities.We decompose the turnaround time into six time periods to find the key points of the process improvement and the influencing factors,and then put forward the reform measures to optimize the blood culture inspection process.MiniTab17.0 statistical software was used to process capability analysis and double sample t test.Results After the implementation of the project,the average turnaround time of the blood culture was shortened from 77.10 h to 64.03 h,improved by 13.06 h(16.94％).Process performance greatly improved in Ppk value increased from 0.49 to 0.88,the benchmark Z value increased from 1.48 to 2.63.After the improvement,except the positive alarm time of blood culture,the mean of the other decomposition time was significantly shorter than before.Conclusions The application of Six Sigma in process management can greatly improve the work efficiency and process performance.This project can save a lot of manpower,material and financial resources,reduce the waiting,shorten turnaround time,that achieve the desired results.

Objective To analyze the expression of urine exosomal miR-375 in prostate tumors and investigate its clinical utility.Methods A total of 45 patients with PCa,24 with benign prostate hyperplasia (BPH) and 24 healthy individuals were enrolled into this study.Exosomes were isolated from the urine of PCa,BPH and healthy individuals and the total RNA was extracted from the exosomes.The exosomal miR-375 expression was assessed by quantitative real-time PCR and analyzed with the comparative quantification cycle method (2-△△CT).We performed comprehensive biostatistical analyses to explore the clinical value of miR-375 in prostate cancer.Results The urine exosomal miR-375 expression was significantly downregulated in the patients with PCa compared with BPH and the healthy controls (P ＜ 0.01).No statistically significant difference of the urine exosomal miR-375 expression levels between the patients with BPH and healthy individuals was observed (P ＞ 0.05).The urine exosomal miR-375 expression level was also found to be associated with clinical stage and bone metastasis status of the patients with PCa (P ＜0.05),and with the increase of Clinical stage.The expression level of miR-375 decreased.No significant relationship was detected between miR-375 level and the patient's age,gleason score and serum prostate-specific antigen level (P ＞ 0.05).Receiver operator characteristic analyses demonstrated that the urine exosomal miR-375 expression could better differentiate PCa from BPH patients:AUC 0.715 (95％ CI:0.589-0.842) vs PSA AUC 0.632 (95％ CI:0.492-0.771) (P＜0.01).Conclusion The urine exosomal miR-375 could serve as a non-invasive biomarker for the diagnosis of PCa.