[How to think about the clinical classifications of acquired aplastic anemia].

Paying attention to the diagnosis and classification of acquired aplastic anemia (AA) is the basis for improving the efficacy and the guarantee for the correct exploration of the pathological mechanism, which is of great clinical and academic significance. At present, AA classification is still based on clinical characteristics, which is a historical product of academic development.It is beneficial to guide symptomatic treatment and for the onset of curative treatment. However, the clinical classification of AA cannot replace the pathological mechanism classification to guide the treatment of the root cause. The classification of the pathological mechanism of AA determines the choice of treatment strategy, and can provide a basis for the study of etiology and prevention, and is also the future research direction. Paying attention to the classification of the pathological mechanism of AA is the basis for improving the efficacy and the guarantee for the correct exploration of the pathological mechanism. Modern medicine has entered the era of "molecular targets" and "precision", and how to treat clinical classification based on clinical characteristics is an important issue faced by clinicians. When many different mechanisms of bone marrow failure isolated from AA patients can be accurately identified, that is, when the clinically diagnosed AA has been truly purified into a disease with a clear pathological mechanism, the clinical classification of AA can help to choose the root cause strategy. This article mainly focuses on how to view the clinical classification of AA for the reference of colleagues.

[Analysis of risk factors for venous thromboembolism in patients with polycythemia vera and establishment of a prediction model].

Objective: To investigate the risk factors of venous thrombosis in patients with polycythemia vera (PV) and establish a prediction model for venous thrombosis. Methods: PV patients with JAK2V617F gene mutation positive in the Second Hospital of Tianjin Medical University from September 2017 to November 2023 were retrospectively included. The patients were divided into groups according to whether they had venous thrombosis. After matching age and gender factors with propensity scores, 102 patients were included in the venous thrombosis group [46 males, 56 females, with a median age M (Q1, Q3) of 52 (44, 60) years] and 204 cases were included in the group without venous thrombosis [92 males, 112 females, with a median age of 52 (44, 59) years]. The clinical and laboratory characteristics, disease progression and incidence of gene mutation were compared between the two groups. The follow-up cohort ended on November 20, 2023, with a median follow-up [M (Q1, Q3)] of 11 (1, 53) years. Multivariate Cox risk model was used to analyze the influencing factors of venous thrombosis in PV patients, and establish a scoring system for the venous thrombosis risk factor prediction model of PV patients. Receiver operating characteristic (ROC) curve was used to evaluate the predictive efficiency of the model. Results: Hemoglobin concentration, the ratio of hematopoietic volume≥55%, neutrophil to lymphocyte ratio≥5, hypertension, subcostal spleen≥5 cm and secondary myelofibrosis in venous thrombosis group were higher than those in non-venous thrombosis group (all P<0.05). In addition, the proportion of history of thromboembolism, V617F gene mutation load (V617F%)≥50%, diabetes mellitus, ASXL1 mutation and secondary reticular silver staining≥3 in the venous thrombosis group were higher than those in the non-venous thrombosis group (all P<0.05). The proportion of PV patients with 3 or more gene mutations was 44.1% (45/102) in venous thrombosis group, which was higher than that of PV patients without venous thrombosis 29.9% (61/204) (P=0.014). The proportion of ASXL1 gene mutation in venous thrombosis group was 17.6% (18/102), which was higher than the 4.9% (10/204) in non-venous thrombosis group (P<0.001). Multivariate Cox risk model analysis showed that previous thromboembolism history (HR=2.031, 95%CI: 1.297-3.179, P=0.002), V617F%≥50% (HR=2.141, 95%CI: 1.370-3.347, P=0.001), ASXL1 mutation (HR=4.632, 95%CI: 1.497-14.336, P=0.008), spleen subcostal≥5 cm (HR=1.771, 95%CI: 1.047-2.996, P=0.033) are the risk factors of venous thrombosis in PV patients. According to HR values, a score system for predicting risk of venous thrombosis in PV patients was established: previous history of thromboembolism, V617F%≥50% and spleen subcostoal≥5 cm were assigned 1 point respectively, and ASXL1 mutation was assigned 2 points. Low risk group: score 0, medium risk group: score 1-2, high risk group: score≥3. The ROC curve analysis of the model for predicting venous thrombosis in PV patients showed that the area under the curve (AUC) was 0.807 (95%CI: 0.755-0.860), with the sensitivity of 88.2% and the specificity of 59.8% when the Youden index was 0.48. Conclusions: Previous thromboembolism history, V617F%≥50%, ASXL1 mutation, spleen subcostoal≥5 cm are risk factors of venous thrombosis in PV patients. The established prediction model has good prediction efficiency.

[Research status and challenges of advanced myeloproliferative neoplasms].

Classical myeloproliferative neoplasms (MPN), also known as Ph-MPN, includes polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). Secondary myelofibrosis (sMF) and secondary acute myeloid leukemia (sAML) are important disease progressions of MPN. After MPN disease progression, hematopoietic stem cells undergo new clonal evolution, leading to drug resistance, poor treatment effect and poor survival of patients. In recent years, the exploration of the mechanism of disease progression and the precise diagnosis and treatment of MPN have attracted much attention. This article summarizes the research status of MPN disease progression, including the pathogenesis, risk stratification, and precision treatment, in order to provide reference for exploring new diagnosis and treatment methods of MPN disease progression.

[The expression and correlation analysis of TOX and inhibitory receptors on peripheral blood CD8+T cells in patients with aplastic anemia].

Objective: To investigate the expression levels of thymocyte selection related high mobility group proteins (TOX) and different inhibitory receptors in peripheral blood CD8+T cells of patients with aplastic anemia (AA), and to conduct correlation analysis. Methods: From September 2019 to November 2020, 27 AA patients in the Department of Hematology, General Hospital of Tianjin Medical University were retrospectively selected, including 21 males and 6 females, with a median age [M (Q1, Q3)] of 48 (30, 72) years. Thirty-three healthy controls, included 17 males and 16 females, with a median age of 46 (27, 69) years. The expression levels of TOX, programmed cell death receptor-1 (PD-1), T-cell immunoglobulin and mucin domain 3 (TIM-3), cytotoxic T-lymphocyte associated antigen-4 (CTLA-4), T-cell immune receptor with immunoglobulin (Ig) and immunoreceptor tyrosine-based inhibitory motif (ITIM) domain (TIGIT), perforin and granzyme B in peripheral blood CD8+T cells from AA patients and healthy controls were detected by flow cytometry. The correlation between TOX expression levels and different inhibitory receptors was analyzed using Pearson correlation analysis. Results: The expression levels of TOX, PD-1, TIM-3, CTLA-4, TIGIT, perforin, and granzyme B in peripheral blood CD8+T cells of AA patients were 47.33%(41.47%, 56.61%), (30.61±12.37)%, (39.94±10.84)%, (6.21±3.40)%, (51.45±20.21)%, (71.32±22.46)%, and (52.39±23.99)%, respectively, which were higher than those of healthy controls 27.32%(21.64%, 46.96%), (21.29±10.01)%, (21.11±3.00)%, (1.31±0.34)% (30.80±13.40)%, (46.72±22.53)%, (21.75±16.43)% (all P<0.05). The expression level of TOX in CD8+T cells was positively correlated with the expression levels of PD-1, TIM-3, CTLA-4, TIGIT, perforin, and granzyme B (r=0.49, 0.65, 0.70, 0.54, 0.58, 0.48, all P<0.05). Conclusion: The expression levels of TOX and different inhibitory receptors on peripheral blood CD8+T cells in AA patients are higher than those in the healthy control group, and the expression levels of TOX and different inhibitory receptors are positively correlated.

[Abnormal expression of CXCR5+CD8+ T cells and CXCL13 in severe aplastic anemia patients and their correlation with hematological parameters].

Objective: To analyze the expression of C-X-C chemokine receptor 5 (CXCR5)+CD8+ T cells and plasma C-X-C motif chemokine 13 (CXCL13) in severe aplastic anemia (SAA) patients and their correlations with hematological parameters. Methods: The clinical data of 35 SAA patients in the Hematology Department of Tianjin Medical University General Hospital from January 2018 to September 2021 were retrospectively analyzed. The patients were divided into two groups according to whether they had received the medication: untreated SAA group and recovery SAA group. In untreated group, there were 18 patients who had not received any medication, with 9 males and 9 females, and aged 51 (18-76) years. In recovery SAA group, there were 17 patients who were separated from component blood transfusion after the immunosuppressive treatment with anti-thymocyte globulin (ATG) combined with cyclosporine A (CsA), with 7 males and 10 females, and aged 46 (16-70) years. Meanwhile, 20 healthy controls were also selected, including 8 males and 12 females, and aged 45(15-72) years. Peripheral blood and bone marrow samples were collected from SAA patients, while peripheral blood samples were obtained from healthy controls. Flow cytometry was used to detect the percentage of CXCR5+CD8+ T cells in peripheral blood and bone marrow samples. The concentration of plasma CXCL13 was measured by enzyme-linked immunosorbent assay (ELISA). The correlations between the percentage of CXCR5+CD8+ T cells and the concentration of CXCL13, as well as the correlations between these two parameters and the hematological parameters were analyzed by Spearman correlation analysis. Results: The proportion of CXCR5+CD8+ T cells in the bone marrow of untreated SAA group was (4.9±2.9)%, which was higher than that of recovery SAA group (2.7±1.5)%, with a statistically significant difference (t=2.34, P=0.027). The proportion of CXCR5+CD8+ T cells in peripheral blood of untreated SAA group, recovery SAA group and healthy control group was (8.4±4.2)%, (3.8±2.3)% and (2.6±2.0)% respectively. The proportion of CXCR5+CD8+ T cells in peripheral blood of untreated SAA group was higher than that of recovery SAA group and healthy control group (both P<0.05). The plasma CXCL13 concentration in untreated SAA group was (97.2±46.8) ng/L, which was significantly higher than that in recovery SAA group [(54.9±20.9) ng/L] and healthy control group [(47.6±17.3) ng/L] (both P<0.05). The proportion of CXCR5+CD8+ T cells in peripheral blood of SAA patients was positively correlated with the concentration of plasma CXCL13 (r=0.545, P<0.001). The proportion of peripheral blood CXCR5+CD8+ T cells in SAA patients was negatively correlated with white blood cell count, platelets count, percentage of neutrophils, absolute neutrophils count, percentage of reticulocytes, absolute reticulocytes count, bone marrow myeloid cells, bone marrow erythroid cells and megakaryocytes count (r=-0.556, -0.392, -0.617, -0.615, -0.395, -0.543, -0.432, -0.484 and -0.523, all P<0.05). The proportion of peripheral blood CXCR5+CD8+ T cells was positively correlated with the percentage of peripheral blood lymphocytes and bone marrow lymphoid cells (r=0.593 and 0.556, both P<0.05). Meanwhile, the concentration of plasma CXCL13 in SAA patients was negatively correlated with white blood cell count, absolute neutrophils count, percentage of reticulocytes, absolute reticulocytes count and bone marrow myeloid cells (r=-0.447, -0.446, -0.498, -0.407 and -0.456, all P<0.05), but positively correlated with bone marrow lymphoid cells (r=0.384, P<0.05). Conclusions: The proportion of CXCR5+CD8+ T cells and the concentration of plasma CXCL13 increases in SAA patients. The proportion of CXCR5+CD8+ T cells in peripheral blood is positively correlated with the concentration of CXCL13. Moreover, the proportion of CXCR5+CD8+ T cells and the concentration of CXCL13 are correlated with many hematological parameters, which may play a critical role in the immune pathogenesis of SAA.

[Clinical and laboratory features of SF3B1-mutated myeloproliferative neoplasms].

Objective: To explore the clinical and laboratory characteristics of SF3B1 gene mutations in myeloproliferative neoplasms (MPN) patients. Methods: The clinical data of 273 MPN patients who were diagnosed MPN and treated in the Second Hospital of Tianjin Medical University from November 2017 to March 2023 were retrospectively analyzed. There were 133 males and 140 females, with a median age M(Q1,Q3)of 56(46, 67) years. The molecular biology and cytogenetic characteristics were detected by second-generation sequencing (NGS) and R+G banding techniques, and the clinical and laboratory characteristics of patients with SF3B1 gene mutation were analyzed. Results: SF3B1 gene mutations were found in 13 patients (4.8%, 13/273).The types of SF3B1 mutations included missense (92.3%, 12/13) and nonsense mutations (7.7%, 1/13).Compared to the non-mutant cohort, patients in SF3B1 mutant cohort had older ages [68(51, 76) vs 56(45, 66)years,P=0.025], higher proportion of splenomegaly [46.2%(6/13) vs 15.8%(41/259),P=0.014]and secondary tumor [23.1%(3/13)vs 3.8%(10/260), P=0.018]with higher proportion of bone marrow blast [0.5%(0, 1.5%) vs 0(0, 0.5%),P=0.002] and lower hemoglobin[(104±36) vs (137±40) g/L,P=0.004] and hematocrit [31%(22%, 40%) vs 41%(35%, 52%),P=0.003]. All of the 10 patients in the SF3B1 mutant cohort whose ring sideroblast (RS) could be evaluated showed no RS formation. The overall survival, thrombosis-free survival and leukemia free survival of MPN patients in SF3B1 mutant cohort were 4.0 (2.0, 6.0), 2.0 (0.5, 4.5) and 4.0 (2.0, 6.0) years, respectively, while patients in the non-mutant cohort were 6.0 (3.0, 10.0), 5.0 (1.0, 8.0), 6.0 (3.0, 10.0) years, respectively, there were no statistical significance between two groups (Z=3.69, 1.66, 2.05, all P>0.05).The secondary tumor free survival of SF3B1 mutant cohort patients was 4.0 (2.0, 6.0) years, which was lower than that of non-mutant cohort patients [5.5 (3.0, 10.0) years, Z=18.18, P<0.001). Conclusions: MPN patients with SF3B1 gene mutations are older, more prone to splenomegaly and secondary tumors. They also have a higher proportion of bone marrow blast, lower hemoglobin and hematocrit, and show no RS formation.

[Efficacy and safety of pegylated interferon alpha-2b for patients with myeloproliferative neoplasm].

Objective: To evaluate the efficacy and safety of pegylated interferon alpha-2b (PEG-IFN-α2b) in the treatment of myeloproliferative neoplasm (MPN). Methods: Thirty-four MPN patients receiving PEG-IFN-α2b treatment in the Second Hospital of Tianjin Medical University from August 2019 to October 2022 were prospectively included. Among the patients, 9 were male and 25 were female, and the median age [M (Q1, Q3)] was 57 (19, 78) years. Patients' clinical characteristics were collected and the follow-up was performed. As of January 30, 2023, the follow-up period [M(Q1, Q3)] was 24 (16, 33) months. The efficacy, safety and changes in immune cell and cytokine levels after 12 and 24 months of treatment were analyzed. Results: During the follow-up period, 4 patients dropped out, and the efficacy was evaluable in 30 patients. Following 12 and 24 months of treatment, the complete hematologic response (CHR) rates were 57.1% (16/28) and 75.0% (18/24), respectively. The complete molecular response (CMR)+partial molecular response (PMR) rates were 27.3% (6/22) and 55.0% (11/20), respectively. The bone marrow histopathological overall response rates (ORR) were 34.6% (9/26) and 47.6% (10/21), respectively. At 12 and 24 months of treatment, the proportions of CD8+HLA-DR+T cells, effector T cell subpopulations, CD56bright natural killer (NK) cells, and plasmacytoid dendritic cells (pDC) were higher than the pre-treatment levels, while the proportion of CD56dim NK cells was lower than the pre-treatment level (all P<0.05). The levels of motif chemokine ligand 10 (CXCL10), tumor necrosis factor (TNF)-α and TNF-ß in bone marrow all increased from those prior to treatment, while the levels of vascular endothelial growth factor (VEGF) and interleukin (IL-4) decreased from those prior to treatment (all P<0.05). Among hematological adverse reactions, white blood cells decrease [47% (16/34)] was observed with high incidence. Among non-hematological adverse reactions, asthenia [44.1% (15/34)] and transaminases increase [32.3% (11/34)] were observed with high incidences. Conclusions: PEG-IFN-α2b has high hematologic, molecular, and bone marrow histopathological response rates in the treatment of MPN. It can reduce malignant clone loads and regulate the immune microenvironment and is safe and well tolerated overall.

[Analysis of risk factors for thromboembolism in patients with JAK2V617F gene mutation positive myeloproliferative neoplasms].

Objective: To analyze the risk factors of thrombosis in patients with JAK2V617F mutation positive myeloproliferative neoplasms (MPN). Methods: A total of 223 MPN patients with JAK2V617F mutation in the Second Hospital of Tianjin Medical University from September 2017 to May 2023 were retrospectively enrolled, including 111 males and 112 females, aged [M(Q1,Q3)] 57(21,66) years. According to the presence or absence of thromboembolism during follow-up, the patients were divided into thrombosis group (n=102) and non-thrombosis group (n=121). The clinical characteristics, laboratory characteristics, cytogenetics and other disease progression and survival of the two groups of patients were analyzed. As of March 31, 2023, the follow-up period [M (Q1, Q3)] was 6 (3, 10) years. The influencing factors of thrombosis in JAK2V617F positive MPN patients were analyzed by using the Cox risk model. Results: Among 223 JAK2V617F positive MPN patients, 144 were polycythemia vera (PV), 51 were essential thrombocythemia (ET) and 28 were primary myelofibrosis (PMF). The mutation rates of ASXL1 and BCORL1 genes in the thrombosis group were 19.6% (20/102) and 6.9% (7/102), respectively, which were higher than those in the non-thrombosis group [9.1% (11/121) and 0.8% (1/121)] (both P<0.05). The proportion of monocytes, C-reactive protein (CRP), interleukin-1ß (IL)-1ß, IL-8 and tumor necrosis factor-ß (TNF-ß) increased in the thrombosis group were higher than those in the non-thrombosis group (all P<0.05). Multivariate analysis showed that age≥60 years (HR=2.132, 95%CI: 1.376-3.303, P=0.001), history of thrombosis (HR=3.636, 95%CI: 2.121-6.202, P<0.001), ASXL1 mutation positive (HR=2.245, 95%CI: 1.093-3.231, P=0.022) and elevated TNF-ß (HR=2.009, 95%CI: 1.113-3.624, P=0.021) were risk factors for thrombosis in JAK2V617F positive MPN patients. Conclusions: In addition to age, history of thrombosis and positive ASXL1 mutation, elevated TNF-ß is also an influencing factor of thrombosis in JAK2V617F positive MPN patients. Intervention of inflammation may have a certain effect on the prevention and treatment of thrombosis.

[Review and perspective of clinical research involving chest tightness variant asthma in China].

Chest tightness variant asthma (CTVA) was first reported and named by Chinese scholars in 2013. It is a new clinical type of asthma characterized by chest tightness as the only or primary symptom, without typical asthma manifestations such as recurrent wheezing and shortness of breath, and without wheezing sounds heard during lung auscultation. The overall epidemiological data on CTVA is currently unavailable. Its pathogenesis is similar to that of typical asthma, involving eosinophilic airway inflammation. Due to the lack of typical clinical manifestations, insufficient knowledge of this disease in some clinicians and some other reasons, CTVA is susceptible to misdiagnosis or missed diagnosis. Currently, the diagnostic criteria for CTVA are: chest tightness as the only or primary symptom, without typical asthma symptoms and signs such as wheezing and shortness of breath, and with any one of the objective indicators of variable airflow limitation. Effective anti-asthma treatment is required, and other diseases that cause chest tightness, such as cardiovascular, digestive, nervous, muscular, and mental diseases should be excluded. CTVA treatment follows that of typical asthma, but the specific treatment duration is uncertain and may require long-term management. Traditional Chinese medicine has shown some therapeutic effects on CTVA. Most CTVA patients have a good prognosis after active anti-asthma treatment. This paper analyzes and summarizes the research of CTVA in China from 2013 and provides new perspectives for further exploration of CTVA.

[Exclusive diagnosis of aplastic anemia].

As an independent disease, aplastic anemia (AA) has been recognized for more than a century. When AA is diagnosed, other non-AA bone marrow failures should be excluded. It is termed as exclusive diagnosis of AA. The exclusive diagnosis of AA is helplessly based on that there is no parameter by which AA can be sensitively and specifically diagnosed now. So further searching for the meaningful diagnostic parameters of AA should be carried on to establish a direct diagnostic protocol of this disease and make it possible to differentiate it clearly from other bone marrow failure disease such as congenenital bone marrow failure, hypoplastic myelodysplastic syndromes, AA-paroxysmal nocturnal hemoglobinuria syndromes, large granules lymphocyte leukemia, clonal cytopenia of undetermined significance, immunorelated pancytopenia, acute hemopoietic arresting and idiopathic cytopenia of undetermined significance. The new markers and technologies being helpful for distinguishing AA from other bone marrow failures should be used in diagnosing AA. Correct understanding and application of exclusive diagnosis is not only related to the correctness of diagnosis and treatment of excluded diseases, but also to the quality of AA diagnosis, treatment and research.

[Effect and prognosis of new type of re-dilated stent in the treatment of pulmonary artery bifurcation stenosis].

the early and med-term follow-up results and technical points of new re-dilated stent in the treatment of pulmonary artery bifurcation opening stenosis, and explore its feasibility and advantages. From March 2019 to October 2020, 10 children [5 males, mean age (7±3) years], mean weight 18.75(13.35,23.05) kg with pulmonary artery bifurcation opening stenosis were treated with new re-dilated stents in the Central China Fuwai Hospital. Including 5 cases of tetralogy of Fallot, 4 cases of pulmonary atresia, 1 case of anomalous origin of coronary artery, all children were given new re-dilated stent implantation. Echocardiography, chest X-ray and electrocardiogram were performed 1 day, 3, 6, 12 months after intervention. Pulmonary artery CTA was performed after 6 or 12 months to evaluate the results, including restenosis, malposition and rupture. A total of 16 stents were implanted in 10 children, 5 cases had simultaneous stenosis of bifurcation openings of pulmonary arteries, and 1 stent was implanted in each of the left and right pulmonary artery openings. The pressure of right ventricular and gradient was significantly decreased immediately after intervention, from preoperative (38-80) mmHg(1 mmHg=0.133 kPa) to postoperative (0-22) mmHg, only one patient's pressure gradient is over 20 mmHg (22 mmHg), and all cases discharged successfully. Stent restenosis, malposition, fracture and other abnormalities were not observed in follow-up. Stents implantation for patients with pulmonary artery bifurcation opening stenosis is very more difficult. A good strategy can ensure that the intervention is safe and effective. It not only avoids the risk of repeated surgery, but also achieves good med-term follow-up results.

[Short- and mid-term outcomes of percutaneous balloon mitral valvuloplasty under the guidance of ultrasound].

Objective: To evaluate the safety, short- and mid-term outcomes of percutaneous balloon mitral valvuloplasty (PBMV) guided by the ultrasound. Methods: In this retrospective study, medical data of 15 patients [9 males and 6 females, with an age of (53±13) years] with PBMV under the guidance of ultrasound in Heart Center of Henan Provincial People's Hospital between December 2016 and January 2019 were collected and reviewed. The short-and mid-term outcomes were analyzed. Results: PBMV was successfully performed in all the patients. One patient underwent surgical valve replacement due to severe mitral regurgitation, and the other 14 patients were all followed up successfully. The average follow-up time was (13.8±4.6) months. Comparisons of preoperative and postoperative data showed significant differences in valve area [(1.84±0.43) cm2 vs (0.89±0.24) cm2], left atrial pressure [(11.9±4.5) mmHg (1 mmHg=0.133 kPa) vs (21.9±6.0) mmHg] and mean mitral valve pressure gradient [(10.9±3.2) mmHg vs (20.1±3.6) mmHg](all P<0.01), with no significant differences in mitral regurgitation area (P=0.67). Postoperative follow-up showed that there were no significant differences in mitral valve area, regurgitation area and N-terminal pro-B-type natriuretic peptide (NT-proBNP) between short-and mid-term postoperatively (all P>0.05). There was no secondary operation due to mitral stenosis in 14 patients, and 3 patients with moderate or severe tricuspid regurgitation showed significant improvement, with gradually recovered cardiac function, and there were no deaths in these patients. Conclusion: PBMV guided by the ultrasound is feasible and effective, and exhibits favorable short-and mid-term outcomes.

[Effects of dynamic contrast enhancement on diffusion weighted imaging score of 3 in prostate imaging reporting and data system version 2 of peripheral zone].

Objective: To investigate the effects and value of dynamic contrast-enhanced (DCE) on diffusion-weighted imaging (DWI) score 3 in the prostate imaging data and reporting system version 2(PI-RADS V2)of peripheral zone. Methods: A retrospective study of consecutive 204 cases of prostate disease in peripheral zone was conducted from January 2015 to January 2018, including 169 cases of prostate cancer and 35 cases of non-prostate cancer. All cases were examined multi-parametric MR imaging on a 3-T MR scanner and confirmed by pathology. Images were analyzed according to PI-RADS V2.Inter-reader agreement of scores was evaluated by kappa coefficient. The constituent ratio of clinically significant prostate cancer (csPCa) in PI-RADS V2 overall scores 3, 3+1 and 4 were calculated and analyzed by non-parametric Chi-square test to determine whether the difference in composition ratio was significant. The positive rate of DCE in diffusion weighted imaging (DWI) score 3 and 4 were calculated. Results: There were 68.6%(140/204) cases in 204 patients with peripheral prostate disease who scored PI-RADS V2 overall score of 3,3+1 and 4. Kappa value was higher for the PI-RADS V2 overallscore of 4 than 3 or 3+1 in the PZ (k values of 0.802 vs 0.737 or 0.591, respectively; all P<0.01). The constituent ratio of csPCa in PI-RADS V2 overallscore of 3, 3+1 and 4 were 16.7%(3/18),43.2%(19/44),78.2%(61/78), the difference of which was significant (χ(2)=29.839, P<0.05).There was also a significant difference between the constituent ratio of csPCa in the score of 3+1 and 4 (χ(2)=15.286, P<0.01).The positive rate of DCE in cases of PI-RADS V2 overall score 3,3+1 and 4 was 76.4%(107/140).The positive rate of DCE incases of DWI score 3 and 4 were 71.0%(44/62), 80.8%(63/78),respectively. Conclusion: DCE has a certain value in PI-RADS V2 due to its high positive rate in the detection of prostate cancer in peripheral zone.DCE is helpful to DWI score 3 to improve the detection rate of csPCa.The PI-RADS V2 overall score 3+1 and 4 in the peripheral zone maybe need to be distinguished due to the different detection of csPCa.

[Lymphocyte activation gene 3 expression on T lymphocyte cell subsets in patients with myelodysplastic syndrome].

Objective: To investigate the expression of Lymphocyte activation gene 3 (LAG3) in myelodysplastic syndromes (MDS) patients. Methods: A total of 16 MDS patients newly diagnosed in Hematology Department of Tianjin Medical University were enrolled from January to November 2019. The healthy control (HC) group includes 16 cases of healthy adults. The expression levels of LAG3 on CD8(+)T cells, CD4(+)T cells and regulatory T cells (Treg) in MDS patients and healthy controls were detected by flow cytometry. Results: A total of 16 patients with MDS were included, including 5 males and 11 females, with a median age of 56 (18-80) years old. HC group includes 16 healthy adults, 8 men and 8 women, with a median age of 40 (17-69) years. There was no statistically significant difference in gender and age composition between the two groups (both P>0.05). The expression of LAG3 on CD8(+)T cells in MDS patients (74.45%±22.31%) was significantly higher than that in HC group (58.78%±14.82%, P<0.05). The LAG3 expression on Treg in MDS patients (64.91%±10.32%) were significantly higher than that of HC group (49.09%±13.58%, P<0.05). There was no statistical difference in LAG3 expression on CD4(+)T cells between the two groups. Conclusion: The expression of LAG3 on CD8(+)T cells and Treg increases in MDS patients than that of healthy people.

[Relationship between SHP-1 gene methylation and STAT3 phosphorylation and prognosis in patients with myelodysplastic syndrome].

Objective: To assess the methylation level of SHP-1 promoter region and the effects on the phosphorylation of the Signal Transducers and Activators of Transcription 3 (STAT3) protein in bone marrow specimen of patients with myelodysplastic syndrome (MDS), and to explore the relationship of SHP-1 methylation and prognosis of the patients. Method: Bone marrow specimens of 93 patients with MDS were collected from the General Hospital of Tianjin Medical University from September 2010 to June 2014. The enrolled subjects included 54 males and 39 females and they were divided into the low-risk group (IPSS score:0-1.0, median: 0.5) and the high-risk group (IPSS score: 1.5-3.0, median: 2.5) according to the International Prognostic Score System (IPSS). The methylation level of SHP-1 was detected by methylation-specific polymerase chain reaction, and the level of p-STAT3 was detected using Western blot. Results: In the high-risk group, 64.44% (29/45) of the patients had methylation in the SHP-1 promoter region, which was significantly higher than the low-risk group 22.92% (11/48). Therefore, SHP-1 methylation was frequently presented in the patients of the high-risk group. Similarly, 66.67% (30/45) of the patients in the high-risk group had positive STAT3 phosphorylation status, whereas only 20.83% (10/48) were tested positive in the low-risk group. In addition, correlation analysis also revealed that the SHP-1 methylation rate was positively correlated with the positive rate of STAT3 phosphorylation (r=0.57,P<0.001). Conclusions: SHP-1 methylation is significantly correlated with the risk of MDS patients. It may be used as an independent predictor of shorter survival in patients of the high-risk group. The increased level of SHP-1 methylation will lead to the uncontrolled activation of the downstream JAK/STAT3 pathway, which in turn can cause further positive feedback to amplify the carcinogenic signal.

[Relationship between anemia and clinical features and prognosis of advanced lung cancer patients].

Objective: To investigate the relationship of anemia and clinicopathological features and prognosis of patients with advanced lung cancer. Methods: The clinical data of 741 patients with stage Ⅲ~Ⅳ lung cancer were collected and analyzed retrospectively. We analyzed the incidence and type of anemia during the natural course of lung cancer patients, and its relationship with the gender, age, duration of disease, clinical stage, prognostic nutritional index (PNI) of these patients. Kaplan-Meier method and multivariate Cox regression model were used to analyze the effect of anemia on prognosis. Results: Among 741 cases of lung cancer patients, 407 (54.9%) cases were accompanied with anemia, whose hemoglobin (Hb) was (89.39±15.76) g/L, including 214 cases of mild anemia, 173 cases of moderate anemia and 20 cases of severe anemia. The most common type of anemia is anemia of chronic disease (ACD), the incidence rate of which was 79.6% (324/407), followed by the iron deficiency anemia (IDA), the incidence rate of which was 4.2% (17/407). The incidence of anemia was marginally related to the gender and age (P>0.05), but significantly related to the duration of disease, clinical stage and PNI (all P<0.05). The degree of anemia was marginally related to the gender and age (P>0.05), but significantly related to the duration of disease, clinical stage and PNI (all P<0.05). The median survival time of the patients with anemia was 10.5 months (95% CI: 10.1~10.9 ), significantly shorter than 13.0 months (95% CI: 12.2~13.8) of patients without anemia (P<0.001). The median survival time of mild anemia patients was 11.0 months (95% CI: 10.7~11.3), significantly longer than 9.6 months (95% CI: 9.1~10.1) of moderate and severe anemia patients (P=0.048). The results of Cox regression survival analysis showed that the incidence and degree of anemia were independent factors of prognosis of patients with lung cancer (P<0.05). Conclusions: During the natural course of advanced lung cancer, the incidence of anemia is high, especially ACD. The incidence and degree of anemia are substantially correlated with the duration of disease, clinical stage and PNI. The incidence and degree of anemia are independent prognostic factors of patients with lung cancer.

[The clinical significance of CD(4)(+)CD(69)(+) T lymphocytes in patients with autoimmune hemolytic anemia/Evans syndrome].

Objective: To investigate the significant of peripheral CD(4)(+) CD(69)(+) T lymphocytes in patients with autoimmune hemolytic anemia (AIHA)/Evans syndrome (ES). Methods: In this study peripheral blood samples from 32 patients with AIHA/ES (15 hemolytic episode patients, 17 remission patients) and 13 healthy controls were collected. Patients with AIHA/ES were recruited in Tianjin Medical University General Hospital from October 2015 to May 2016. The percentages of CD(69)(+) T lymphocytes were analyzed by flow cytometry. The expression of CD(69) mRNA in CD(4)(+) T lymphocytes which was sorted from peripheral blood by magnetic activated cell sorting (MACS) was detected using real-time PCR. Soluable CD(69) was measured by ELISA. Results: In hemolytic episode patients, the ratio of CD(3)(+)CD(69)(+)/CD(3)(+)T lymphocytes [(3.08±1.48)%] was significantly higher than that in healthy controls [(1.28±0.83)%, P<0.01] and in remission group[(1.96±1.33)%, P<0.05]. The absolute count of CD(3)(+)CD(69)(+)T lymphocytes in hemolytic episode group [(2.94±1.81)×10(7)/L] was higher than that in healthy controls [(1.48±1.42)×10(7)/L, P<0.05]. The ratio of CD(3)(+)CD(4)(+)CD(69)(+)/CD(3)(+)CD(4)(+)T cells in hemolytic episode group [(2.16±1.56)%] was significantly higher than that in remission group [(1.16±0.62)%, P<0.05] and healthy controls[(0.94±0.78)%, P<0.05]. The quantity of CD(3)(+)CD(4)(+)CD(69)(+)T lymphocytes in hemolytic episode group[(1.04±0.98)×10(7)/L] was higher than in healthy controls [(0.44±0.38)×10(7)/L, P<0.05]. The ratio of CD(3)(+)CD(8)(+)CD(69)(+)/CD(3)(+)CD(8)(+)T lymphocyte in hemolytic episode group [(4.87±2.56)%] was significantly higher than that in healthy controls[(1.83±1.27)%, P<0.01]. The quantity of CD(3)(+)CD(8)(+)CD(69)(+)T lymphocytes in three groups did not show significant difference. The ratio of CD(3)(+)CD(4)(+)CD(69)(+)/CD(3)(+)CD(4)(+) T lymphocytes in hemolytic episode group was negatively correlated with hemoglobin (Hb) (P<0.01) , positively correlated with the percentage of reticulocytes (Ret%) (P=0.01) total bilirubin(TBil), indirect bilirubin(IBil) (P<0.01) and not correlated with absolute reticulocytes count, lactic dehydrogenase (LDH), complement 3(C3), complement 4 (C4). The ratio of CD(3)(+)CD(4)(+)CD(69)(+)/CD(3)(+)CD(4)(+)T lymphocytes in remission group was negatively correlated with Hb (P<0.05). In hemolytic episode patients CD(69) mRNA (32.26±35.11) was significantly higher than that in remission group(6.05±5.87) (P<0.05) and healthy controls (1.76±1.85)(P<0.01). CD(69) mRNA in remission group was significantly higher than healthy controls (P<0.05). Serum CD(69) in hemolytic episode patients [(494.21±16.06) ng/L] was significantly higher than that in healthy controls [(441.39±104.6) ng/L, P<0.05]. Conclusion: Our findings suggest that the proportion of CD(4)(+)CD(69)(+) T lymphocytes increase in AIHA/ES patients, which is correlated with the severity of disease.

[Significance of anti-EPO receptor antibody in immune-related pancytopenia].

Objective: To confirm the presence of erythropoietin receptor (EPOR) antibody in patients with immune-related pancytopenia (IRP) and to evaluate the significance of EPOR in IRP. Methods: A total of 59 newly-diagnosed IRP patients, 62 patients with IRP in remission, 14 patients with aplastic anemia (AA), 15 patients with myelodysplastic syndromes (MDS) and 33 healthy controls were enrolled in this study from January 2013 to June 2015 in Tianjin Medical University General Hospital. The anti-EPOR antibody was detected by enzyme-linked immunosorbent assay(ELISA). The expression of EPOR mRNA was detected by quantitative real-time PCR (qRT-PCR). The correlation between the data and the clinical indicators of patients was analyzed. Results: The levels of anti-EPOR antibodies were higher in the newly-diagnosed IRP patients than in the remission IRP group, AA group, MDS group and controls(0.84±0.39 vs 0.46±0.25, 0.49±0.25, 0.50±0.25, 0.53±0.14, all P<0.05). The expression of EPOR- mRNA was up-regulated in the newly-diagnosed IRP group than in the remission IRP group and the controls. The positive rate of EPOR antibody was significantly elevated in the patients with low hemoglobin level (<100 g/L), positive GlycoA antibody, or low complement C3 (all P<0.05). Anti-EPOR antibody was significantly decreased in the IRP patients responding to immunosuppressive therapy. Conclusions: Anti-EPOR autoantibody is present in patients with IRP. Detection of anti-EPOR antibody has important clinical value in the diagnosis, differential diagnosis and treatment effect evaluation of IRP.

[Effects of different computed high b-values on diffusion weighted imaging scores in Prostate Imaging Reporting and Data System version 2 of prostate cancer in peripheral zone].

Objective: To determine the effects of different computed high b-value on diffusion weighted imaging (DWI) scores in Prostate Imaging Reporting and Data System version 2 (PI-RADS v2) of prostate cancer in peripheral zone. Methods: A retrospective study of 104 cases of prostate cancer in peripheral zone was conducted, all of the patients were histopathologically confirmed by transrectal ultrasound guided saturation biopsy or radical prostatectomy in Tongji Hospital of Tongji University from January 2012 to December 2015.All MRI imaging examinations were performed by using a 3.0T Siemens Verio MRI scanner.The imaging protocol consisted of high-resolution axial and sagittal T(2) weighted imaging (T(2)WI), axial acquired diffusion weighted imaging (aDWI) with b=0, 50, 1 000 s/mm(2) and dynamic contrast-enhanced (DCE) scans.Computed diffusion weighted imaging (cDWI) images with b=1 000, 1 400, 2 000 s/mm(2) were processed by Matlab.These three groups of cDWI images were analyzed according to the PI-RADS v2 criteria, and signal intensity of ratio (SIR) of lesions were analyzed by independent t test and one-way ANOVA in each group. Results: The numbers of cases with a DWI score of 2 and 5 were similar among three groups.Nine cases (33.3%) in all 27 cases with a DWI score of 3 on b=1 000 s/mm(2) upgraded to score 4 when b-value rose to 1 400 and 2 000 s/mm(2).The ratios of SIR of lesions in cases upgraded from DWI score 3 to 4 to those unchanged cases on b=1 400 and 2 000 s/mm(2) were 1.86±0.21 to 1.61±0.27 and 2.18±0.26 to 1.75±0.30, respectively (t=2.486, t=3.671, both P<0.05). In these 9 cases who upgraded to DWI score 4, SIRs of the lesion were significantly different between groups when b=1 000, 1 400 and 2 000 s/mm(2) (F=10.907, 33.768, 8.043, all P<0.05), and their SIRs increased with the rising of b-value. Conclusions: The computed high b-value (b≥1 400 s/mm(2)) mainly affects cases with a DWI score of 3, but DWI scores would not change neither in b=1 400 s/mm(2) nor in 2 000 s/mm(2) cases. For DWI score in PI-RADS v2 of prostate cancer in peripheral zone, b=1 400 s/mm(2) is probably of more scoring value than b=1 000 or 2 000 s/mm(2).

[Quantities and function of NK cells in patients with immune thrombocytopenia].

Objective: To investigate natural killer (NK) cell quantities and function in patients with immune thrombocytopenia (ITP) . Methods: A total of 66 ITP patients (34 newly diagnosed and 32 in complete remission) were collected from September 2015 to May 2016 in Tianjin Medical University General Hospital, and 30 healthy volunteers were recruited as controls. The percentages of NK cells and their subsets in peripheral blood, the expression of activating receptor (NKp44), inhibitory receptor (NKG2A) and CD16, perforin and granzyme ß were detected by flow cytometry. The correlation between the above parameters and patients' immune status and platelet level were evaluated. Results: (1)The percentage of CD3(-)CD56(+) NK cells in newly diagnosed patients (10.99%±4.89%)and patients in complete remission (9.73%±6.75%) were significantly lower than that in healthy controls (14.67%±7.24%)(P=0.023, 0.003). The percentage of NK cells Bright subset was significantly lower in the newly diagnosed patients(0.48%±0.23%)and those in complete remission (0.41%±0.33%) than in healthy controls(0.64%±0.32%)(P=0.037, 0.002); the percentage of Dim subset was also significantly lower in the newly diagnosed (10.16%±5.02%) and patients in complete remission (8.07%±5.74%) than in healthy controls(14.16%±7.19%) (P=0.009, 0.007). (2)The proportion of Bright subset in total NK cells in new diagnosed ITP patients (6.48%±4.33%) was significantly higher than that in healthy controls (4.21%±2.70%)(P=0.020); the proportion of Dim NK cells subset in new diagnosed ITP patients (93.51%±4.33%) was significantly lower than that in healthy controls(95.79%±2.70%) (P=0.020). (3)The expression of activating receptor NKp44 in new diagnosed ITP patients was significantly lower than that in complete remission group and healthy controls[0.28%(0.95%)vs 0.61%(2.05%), 0.92%(0.90%); P=0.047, 0.048]; the expression of inhibitory receptor NKG2A in new diagnosed ITP patients was significantly higher than that in healthy controls(42.34%±23.86% vs 29.25%±12.83%, P=0.009). The proportion of CD16 was significantly lower in the newly diagnosed patients than in healthy controls(93.51%±4.33%95.79%±2.70%, P=0.020). (4)The expression of perforin in the newly diagnosed ITP patients was significantly lower than that in healthy controls [87.52%(25.29%)vs 91.55%(8.29%), P=0.025]; the expression of granzyme ß in ITP patients and controls showed no statistically significant difference. (5)The level of NK cells in ITP patients was negatively correlated with CD3(+) CD8(+) T cells (r=-0.387, P=0.012) and CD5(+) CD19(+) B cells in peripheral blood (r=-0.273, P=0.028), positively correlated with the ratio of CD3(+) CD4(+) /CD3(+) CD8(+) (r=0.358, P=0.028) and peripheral platelet count (r=0.314, P=0.011). Conclusion: Deceased quantities and impaired total NK function, insufficient suppression of autoreactive T and B cells might play a role in the pathogenesis of ITP.