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 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: 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 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 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 investigate the molecular epidemiological characteristics and homology of Staphylococcus aureus (S. aureus)isolated from patients in intensive care units (ICUs)of a hospital,so as to provide laboratory basis for the effective control of healthcare-associated infection(HAI). Methods 62 S. aureus strains isolated from various specimens from ICU patients with infection in March-August 2013 were collected,7 housekeeping genes were amplified with polymerase chain reaction (PCR),the amplified products were sequenced,ST typing of strains was performed by multilocus sequence typing (MLST ), phylogenetic analysis of ST typing was conducted. Results 62S. aureus strains were amplified specific product of 7 housekeeping genes;there were 10 ST genotypes, in which 2 ST genotypes(STn1and STn2)were first discovered,1 ST genotype(ST675)was first discovered domes-tically. ST239 was the main ST type of S. aureus from ICU patients in this hospital,accounting for 74.20% ,which distributed in 6 ICUs,ST5 distributed in 3 ICUs. 62 strains formed 7 main branches in the phylogenetic tree,55 (88.71% )MRSA strains were detected. Conclusion S. aureus isolated from hospital ICUs has some homology, and the small number of types showed the trend of concentrated distribution.

Objective To investigate the changes of peripheral blood tacrolimus concentration and lymphocyte subsets in the uterus transplant recipient,and provide the evidence for monitoring the immune status after uterus transplantation.Methods The peripheral blood tacrolimus concentrations of the uterus transplant recipient during 1 year after transplantation were measured with the microparticle enzyme immunoassay (MEIA).Meanwhile,the whole blood cell counts and lymphocyte subsets were determined by the blood analyzer and flow cytometer,respectively.Results The blood tacrolimus concentrations of the uterus transplant recipient in the first month and second month after transplantation were (13.51 ± 3.92) ng/mL and (15.58 ± 1.19) ng/mL,respectively.The lymphocyte absolute counts were normal before transplantation.At the fifth day after transplantation,the counts of CD3 + T lymphocytes,CD4 + T lymphocytes,CD8 + T lymphocytes and NK cells and the ratio of CD4/CD8 were significantly decreased.One week after transplantation,the counts of CD4 + T lymphocytes were recovered to the normal range and maintained,but its recovery was slower than that of CD8 + T lymphocytes.The ratio of CD4/CD8 ranged from 0.4 to 0.8 during 10 days after transplantation,and increased and maintained between 0.8 and 1.1 after that.The counts of NK cells increased gradually from the 10th day after transplantation,but still did not recover to the level before transplantation even at the 20th day after transplantation.However,the counts and percentages of B lymphocytes did not decrease but increased at the fifth day after transplantation,and recovered to normal gradually from the 10th day after transplantation.There was no significant correlation between the CD3 + T lymphocyte count and blood tacrolimus concentration.Conclusion The dynamic changes of blood lymphocyte subsets and tacrolimus concentration exist in the uterus transplant recipient,which need to be further verified by a large amount of clinical data.