Objective:To explore the standardized performance of a FISH probe before clinical detection.Methods:The probe sensitivity and specificity of ETV6/RUNX1 were analyzed via interphase and metaphase FISH in 20 discarded healthy bone marrow samples. The threshold system of the probe was established using an inverse beta distribution, and an interpretation standard was established. Finally, a parallel-controlled polymerase chain reaction detection study was conducted on 286 bone marrow samples from patients at our hospital. The clinical sensitivity, specificity, and diagnostic coincidence rate of ETV6/RUNX1 FISH detection were analyzed, and the diagnostic consistency of the two methods was analyzed by the kappa test.Results:The probe sensitivity and specificity of the ETV6/RUNX1 probe were 98.47% and 100%, respectively. When 50, 100, and 200 cells were counted, the typical positive signal pattern cutoffs were 5.81%, 2.95%, and 1.49%, respectively, and the atypical positive signal pattern cutoffs were 13.98%, 9.75%, and 6.26%, respectively. The clinical sensitivity of FISH was 96.1%, clinical specificity was 99.6%, diagnostic coincidence rate was 99.00%, diagnostic consistency test kappa value was 0.964, and P value was <0.001.Conclusion:For FISH probes without a national medical device registration certificate, standardized performance verification and methodology performance verification can be performed using laboratory developed test verification standards to ensure a reliable and accurate reference basis for clinical diagnosis and treatment.

Background::Gastric cancer (GC), a malignant tumor with poor prognosis, is one of the leading causes of cancer-related deaths worldwide; consequently, identifying novel therapeutic targets is crucial for its corresponding treatment. NUF2, a component of the NDC80 kinetochore complex, promotes cancer progression in multiple malignancies. Therefore, this study aimed to explore the potential of NUF2 as a therapeutic target to inhibit GC progression. Methods::Clinical samples were obtained from patients who underwent radical resection of GC at Lanzhou University Second Hospital from 2016 to 2021. Cell count assays, colony formation assays, and cell-derived xenotransplantation (CDX) models were used to determine the effects of NUF2 on GC progression. Flow cytometry was used to detect the effect of NUF2 or quercetin on cell cycle progression and apoptosis. A live-cell time-lapse imaging assay was performed to determine the effect of NUF2 on the regulation of mitotic progression. Transcriptomics was used to investigate the NUF2-associated molecular mechanisms. Virtual docking and microscale thermophoresis were used to identify NUF2 inhibitors. Finally, CDX, organoid, and patient-derived xenograft (PDX) models were used to examine the efficacy of the NUF2 inhibitor in GC. Results::NUF2 expression was significantly increased in GC and was negatively correlated with prognosis. The deletion of NUF2 suppressed GC progression both in vivo and in vitro. NUF2 significantly regulated the mitogen-activated protein kinase (MAPK) pathway, promoted G2/M phase transition, and inhibited apoptosis in GC cells. Additionally, quercetin was identified as a selective NUF2 inhibitor with low toxicity that significantly suppressed tumor growth in GC cells, organoids, CDX, and PDX models. Conclusions::Collectively, NUF2-mediated G2/M phase transition and apoptosis inhibition promoted GC progression; additionally, NUF2 inhibitors exhibited potent anti-GC activity. This study provides a new strategy for targeting NUF2 to suppress GC progression in clinical settings.

Objective:To establish and validate analytic performance of laboratory-developed real-time quantitative polymerase chain reaction (RQ-PCR) test (LDT) for BCR::ABL1 p210 transcript.Methods:Using the primes and probes released by the Europe Against Cancer Program(EAC), we have established BCR::ABL1 p210 RQ-PCR test. The laboratory-specific conversion factor (CF) was determined by the WHO 1 st International Genetic Reference Panel, and a two-level internal control is developed using a mixture of K562 and HL60 cell lines was created to ensure traceability. Analytical performance, including analytical accuracy, analytical precision, linearity range, analytic sensitivity and specificity of RQ-PCR LDT test for BCR::ABL1 p210 transcript were validated according to CLIS guidelines. Furthermore, a comparison was made with an FDA-cleared RQ-PCR in vitro diagnostics (IVD) kit by Bland-Altman analysis. Results:The laboratory specific conversion factor (CF) for LDT RQ-PCR was determined to be 0.535 based on WHO 1 st International Genetic Reference Panel, which can be used to convert to the BCR::ABL international scale (BCR::ABL1 IS) reliably. The repeatability of BCR::ABL1 IS results at 4 different molecular response (MR0.5,MR1.5,MR2.5,MR3.5) levels are 7.44%, 5.33%, 9.12% and 18.06%, respectively, with total precision of 7.99%, 5.49%, 10.95% and 17.99%. The previous CAP proficiency test (PT) results from our laboratory were within the acceptable range of variation. MR results of our laboratory and MR mean value of all CAP-PT laboratory is highly correlated ( r=0.999, P<0.01), and consistent according to Bland-Altman analysis. Furthermore, the LDT method in our laboratory has a high correlation with the test results of FDA-cleared Qiagen IVD kit ( r=0.997, P<0.01). BCR::ABL1 IS results of BCR::ABL1 e13a2 transcript showed linearity within the range of 0.001%-7.454%, with a maximum coefficient of variation (% CV) 64.09%. The linearity range of e14a2 transcript BCR::ABL1 IS was 0.002%-12.398%, with a maximum % CV of 43.37%. The test has a limit of detection (LoD) of MR5.0 (0.001% IS) for e13a2 and MR4.8 (0.002% IS) for e14a2 transcript, respectively. The limit of quantitation (LoQ) for both e13a2 and e14a2 transcripts was MR4.7 (0.002% IS). The test exhibited 100% specificity, with no cross-reactivity observed between the p190 transcript and p210. Conclusions:The analytic performance of BCR::ABL1 p210 LDT RQ-PCR test from our laboratory is excellent, which can meet the clinical needs of BCR::ABL1 detection in patients with chronic myeloid leukemia.

Objective To explore the feasibility of quartz glass for radiotherapy dosimetry through the experimental study of the thermoluminescence characteristics of synthetic quartz glass. Methods The thermoluminescence glow curves of quartz glass under different annealing conditions were analyzed, the thermoluminescence characteristics of quartz glass were studied, and the measurement parameters were optimized. Using the Co-60 reference radiation field in the National Secondary Standard Dosimetry Laboratory, the quartz glass samples under different annealing conditions were irradiated following the dose levels of radiotherapy, i.e., 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 10.0 Gy, respectively. According to the relationship between the absorbed dose of quartz glass and the relative thermoluminescence signal intensity, the linearity and dispersion of the dose response of quartz glass were obtained, and the feasibility of quartz glass for radiotherapy dosimetry was analyzed. Results The linear correlation coefficient of dose response of quartz glass under annealing condition of 430℃ for 10 min was 0.9984, and the dose response dispersion was 0.97% at the absorbed dose of 2 Gy. The linear correlation coefficient of dose response of quartz glass under annealing condition of 600℃ for 1 h was 0.9911, and the dose response dispersion was 1.4% at the absorbed dose of 2 Gy. Conclusion Preliminary results suggest that quartz glass with annealing condition of 430℃ for 10 min has the potential to be used for radiotherapy dosimetry.

Objective:To observe the therapeutic effect of quercetin (QUE) on triggering receptor expressed on myeloid cells (TREM-1) activated macrophage inflammation and lipopolysaccharide (LPS) induced acute lung injury (ALI) in mice, and explore its possible mechanism.Methods:In vitro cell experiment: The primary peritoneal macrophages of mice were collected by intraperitoneal injection of 3% calcium mercaptan acetate. The collected cells were divided into blank control group, dimethylsulfoxide (DMSO) vehicle group, TREM-1 agonist group (10 μg/ml), QUE group (10 μmol/L) and TREM-1 agonist + QUE group (cells were pretreated with 10 μmol/L QUE for 30 min before adding agonist). Enzyme linked immunosorbent assay (ELISA) was used to detect the secretion of interleukin (IL)-1β, tumor necrosis factor (TNF)-α and IL-6 in the culture supernatant of primary macrophages; To observe the effect of QUE on LPS-induced TREM-1 protein levels, macrophages were divided into: normal control group, LPS group (100 ng/ml) and LPS+ QUE treatment group [macrophages were pretreated with 10 μmol/L QUE for 2 hours, and then incubated with LPS (100 ng/ml) for 16 hours]. Western blot was used to detect the expression of TREM-1 protein. In animal experiments: 80 male C57BL/6 mice were randomly divided into 4 groups (20 in each group): normal control group, ALI model group, QUE group and QUE treatment group (LPS+ QUE). In the ALI model group, the ALI model was established by intratracheal injection of 5 mg/kg LPS; The mouse ALI model was established by intratracheal injection of LPS 5 mg/kg in the QUE treatment group, and then intraperitoneal injection of 15 mg/kg QUE. The control group was given the same amount of normal saline intratracheal followed by intraperitoneal injection of DMSO, and the QUE group was given the same amount of normal saline intratracheal followed by intraperitoneal injection of 15 mg/kg QUE. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissue in each group; Inflammatory cells including IL-1β, TNF- α and IL-6 in bronchoalveolar lavage fluid (BLAF) of mice in each group were counted ; The expression of TREM-1 mRNA and protein in lung tissue of mice in each group was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot. Results:In vitro cell experiment: the secretion of IL-1β, TNF-α and IL-6 in the supernatant of primary macrophages in TREM-1 agonist group was higher than those in DMSO vehicle group, while the secretion of IL-1β, TNF-αand IL-6 in the supernatant of primary macrophages in TREM-1 agonist + QUE group were lower than that of TREM-1 agonist group (all P<0.001). The expression of TREM-1 protein in LPS group was higher than that in control group ( P<0.05), while the expression of TREM-1 protein in LPS + QUE group was lower than that in LPS group ( P<0.05). Animal experiments showed that compared with the control group, the ALI model group had higher lung pathological injury score, more total cells, macrophages and neutrophils in BALF and increased TNF-α, IL-6, IL-1β content (all P<0.001). The above indexes in QUE group were lower than those in ALI model group (all P<0.001). The results of qRT-PCR and Western blot showed that compared with the control group, the expression of TREM-1 mRNA and protein in the lung tissue of ALI model group was increased, while the expression of TREM-1 mRNA and protein in the lung tissue of QUE group was lower than that of ALI model group (all P<0.05). Conclusions:Quercetin can inhibit TREM-1 activation, reduce macrophage inflammatory response and LPS induced acute lung injury in mice.

Acute respiratory distress syndrome (ARDS) is a common cause of critical illness and high mortality from respiratory failure. Increased dead space fraction (VD/VT) was independently associated with lung injury and mortality of ARDS. VD/VT is readily obtained by bedside measurements of arterial blood gas and end-tidal carbon dioxide. Early attention and application of VD/VT as an indicator will help to better understand the pathophysiological of ARDS, guide clinical treatment, and better assess the severity and clinical prognosis of the disease.

Objective:To retrospectively analyze the clinical and serological characteristics in rehabilitated patients with common novel coronavirus pneumonia(COVID-19).Methods:A total of 165 patients with common COVID-19 were enrolled in this retrospective study, in which clinical data was collected from February 23 to March 15, 2020 in Leishenshan Hospital(Wuhan, China). The patients with COVID-19 were divided into elderly group and non-elderly group according to their age, and the differences in the clinical and serological metabolic characteristics between these two groups were analyzed.Results:49.7% patients were over 60 years old. The most common clinical symptoms were fever, cough, and fatigue, followed by muscle soreness. Expectoration and digestive tract symptoms were rare. Dyspnea occurred more frequently in the elderly group than in non-elderly group(47.56% vs 25.30%, P<0.01). Hypertension was the most common concomitant disease(accounting for 29.1%)followed by diabetes. Hypertension was more common in the elderly group than in non-elderly group(41.46% vs 16.86%, P<0.01), but without significant difference in diabetes between the two groups. The counts of leukocytes and lymphocytes in all patients were in the normal range, and no difference was observed between the groups. The comparison of serological indicators showed that serum creatinine in the elderly group was higher than that in the non-elderly group( P<0.01)while serum albumin, glomerular filtration rate, and serum calcium were lower in the elderly group. After serum albumin correction, the levels of albumin corrected calcium in all patients were in the normal range, without significant difference between these two groups. There was no significant difference between the two groups when the length of hospital stay was taken as the index of outcome [(34.01±10.24) vs(30.97±10.51)d, P>0.05]. Conclusion:Fever, cough, and fatigue are the most common clinical symptoms in patients with ordinary COVID-19. The elderly are more likely to develop dyspnea. The blood routine and metabolic characteristics in patients with common COVID-19 are normal, but serum albumin level is more likely to decrease in elderly patients with COVID-19.

Objective:To explore predictors of overall survival (OS) in Chinese patients with polycythemia vera (PV) .Methods:A total of 906 consecutive newly diagnosed patients with PV seen at the Blood Diseases Hospital, Chinese Academy of Medical Sciences, from June 2007 to February 2020 were included, and their data were collected. PV was diagnosed according to 2016 World Health Organization (WHO) diagnostic definitions. OS and prognostic factors were retrospectively analyzed.Results:Among the 906 patients, 439 were male (48.5%) and 467 were female (51.5%) . The median age was 57 years (range: 18-91 years) . 31.6% (276/874) of the patients had a thrombosis history at diagnosis, and 4.6% (25/541) of the patients had abnormal cytogenetics. The median follow-up was 54 months (95% confidence interval [ CI] 8-130 months) . The 5- and 10-year cumulative deaths were 5.8% (95% CI 4.8%-6.7%) and 11.1% (95% CI 9.3%-12.9%) , respectively. Univariate analysis showed that age ≥60 years, thrombosis history, white blood cells (WBC) ≥15×10 9/L, platelet (PLT) ≥450×10 9/L, and platelet distribution width (PDW) ≥15 fl significantly correlated with worse OS, and palpable spleen correlated with better OS. Multivariate analysis showed that age ≥60 years ( HR=4.3, 95% CI 2.1-9.2, P<0.001) and PDW ≥15 fl ( HR=2.1, 95% CI 1.1-4.0, P=0.023) were independent prognostic factors for worse OS. The 5-year cumulative death for patients with PDW ≥15 fl or PDW<15 fl was 8.6% (95% CI 5.9%-11.3%) or 4.4% (95% CI 3.4%-5.4%) , respectively. The 5-year cumulative death for patients defined as low-, intermediate-, and high-risk patients by international working group score system for PV (IWG-PV) were 0.8% (95 CI 0.2%-1.4%) , 4.0% (95% CI 2.7%-5.3%) , and 12% (95% CI 9.6%-14.4%) , respectively, with a significant difference among the three cohorts ( P<0.05) . PDW ≥ 15 fl significantly affected OS for intermediate- and high-risk patients ( HR=2.3, 95% CI 1.2-4.2, P=0.009) defined by IWG-PV score system, but not for low-risk patients ( HR=3.1, 95% CI 0.2-52.0, P=0.405) . Conclusions:Age ≥60 years and PDW ≥15 fl were independent prognostic factors for worse OS in PV. IWG-PV score system effectively predicted OS for Chinese patients with PV.

Chronic myeloid leukemia (CML) has made a milestone progress due to the development of the first generation tyrosine kinase inhibitor(TKI). Nowadays, most clinical trials in CML focus on discontinuation, even the second discontinuation, and the third generation TKI against T315I mutation. The conventional treatments are more focused on decreasing BCR-ABL transcripts rapidly. At the same time, the treatment management of some special patients has been valued.

Objective To establish a reliable pretreatment method for the detection of mequindox and its metabolite in pork luncheon meat by ultra-performance liquid chromatography/triple qudrupole tandem mass spectrometry. Methods Samples were extracted with ethyl acetate, and the results of purification and enrichment by PAX and PEP solid-phase extraction columns were analyzed. Acetonitrile/methanol (3:11) - 0.1% formic acid water was used as the mobile phase, and Shimadzu Inertsil ODS-3-column (3µm, 2.1 × 100mm) chromatographic columns were used for qualitative and quantitative analysis using the multi-reaction detection positive ion mode. Results The results showed that PEP cartridge had good recovery rate. The detection limit of mequindox was 0.10µg/kg, and limit of quantitation was 0.30µg/kg. The average recoveries for spiked levels of 0.33, 0.83, and 1.65µg/kg were 127%, 72.0%, and 60.1%, respectively. The detection limit of 2-quinoxalinecarboxylic acid was 0.10µg/kg, and limit of quantitation was 0.40µg/kg. The average recoveries for spiked levels of 0.42, 1.05, and 2.1µg/kg were 125%, 99.0%, and 60.9%, respectively. Conclusion This method is suitable for the determination of mequindox and its metabolite 2-quinoxalinecarboxylic acid in luncheon meat.