Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal hematologic disorder. Historically, most PNH patients mainly received supportive treatment, which led to poor quality of life and high mortality rates. The advent of downstream complement C5 inhibitors has dramatically changed the natural course of PNH. These inhibitors alleviate clinical symptoms, significantly reduce severe complications, and improve survival rates. Despite their ability to control intravascular hemolysis and enhance hemoglobin levels, approximately half of the treated patients still require blood transfusions due to extravascular hemolysis (EVH) mediated by upstream complement component C3. Upstream complement inhibitors not only control IVH but also reduce the deposition of C3 fragments, thereby solving the problem of EVH and further improving the clinical outcomes of PNH patients. However, due to the cascade of complement activation and the increased accumulation of PNH red blood cells after treatment with upstream complement inhibitors, there may be an increased risk of more severe breakthrough hemolysis events. Additionally, the potential risk of infections associated with complement inhibitors is another issue that needs to be addressed.

Objective:To establish a rapid method for urinary iodine determination based on arsenic-cerium catalytic spectrophotometry using semiconductor water bath rapid cooling digestion technology (rapid cooling method) and an automated iodine analyzer.Methods:Urine samples were collected from three staff members randomly selected at the Gansu Provincial Center for Disease Control and Prevention. The samples were mixed and divided into three equal portions. Using rapid cooling method instead of traditional natural cooling method, the mixed urine sample was lowered from 100 ℃ to the measurement temperature (30 ℃), and then the urine iodine was determined by arsenic cerium catalytic spectrophotometry using a fully automatic iodine analyzer. The differences between the two cooling methods were compared. Additionally, the rapid cooling-arsenic-cerium catalytic spectrophotometry method was validated in terms of the linear equation of the standard curve, precision, accuracy (using urinary iodine reference materials GBW09110e and GBW09109m, and spike recovery experiments), and detection limit.Results:Using the rapid cooling method, the temperature of urine samples reached 30 ℃ in 40 minutes, while the natural cooling method required 120 minutes to reach the same temperature. Urinary iodine levels determined by the rapid cooling method in the three samples were (158.3 ± 1.9), (133.7 ± 2.7), and (219.2 ± 3.1) μg/L, respectively, while those determined by the natural cooling method were (155.5 ± 2.7), (136.2 ± 2.3), and (215.1 ± 3.9) μg/L, respectively. There was no statistically significant difference between the results of the two methods ( t = 1.43, P = 0.192). In the validation experiments for the rapid cooling method, the linear equations for the low-concentration (0 - 300 μg/L) and high-concentration (300 - 1 200 μg/L) iodine standard series were y = - 0.003 0 x + 0.117 5 ( r = - 0.999 7) and y = - 0.001 5 x + 0.649 3 ( r = - 0.999 8), respectively. The precision experiments showed that the relative deviation values of the repeated tests for two different iodine concentrations (300 and 600 μg/L) were 1.99% and 1.90%, respectively. The accuracy experiments demonstrated that the repeated test results for the national urinary iodine reference materials (GBW09110e and GBW09109m) were 216.8 and 134.7 μg/L, respectively, with relative standard deviations of 1.04% and 1.35%, respectively, all within the reference ranges[(218 ± 15) and (135 ± 10) μg/L]. The detection limit was 1.20 μg/L. The spike recovery rate ranged from 92.90% to 99.10%, with an average recovery rate of 96.77%. Conclusions:The rapid cooling-arsenic-cerium catalytic spectrophotometry method not only shortens the time required for urinary iodine determination and improves the speed of measurement, but also provides accurate results, which can be applied when a large number of urine samples need to be measured in a short period of time.

Objective:To establish a rapid method for urinary iodine determination based on arsenic-cerium catalytic spectrophotometry using semiconductor water bath rapid cooling digestion technology (rapid cooling method) and an automated iodine analyzer.Methods:Urine samples were collected from three staff members randomly selected at the Gansu Provincial Center for Disease Control and Prevention. The samples were mixed and divided into three equal portions. Using rapid cooling method instead of traditional natural cooling method, the mixed urine sample was lowered from 100 ℃ to the measurement temperature (30 ℃), and then the urine iodine was determined by arsenic cerium catalytic spectrophotometry using a fully automatic iodine analyzer. The differences between the two cooling methods were compared. Additionally, the rapid cooling-arsenic-cerium catalytic spectrophotometry method was validated in terms of the linear equation of the standard curve, precision, accuracy (using urinary iodine reference materials GBW09110e and GBW09109m, and spike recovery experiments), and detection limit.Results:Using the rapid cooling method, the temperature of urine samples reached 30 ℃ in 40 minutes, while the natural cooling method required 120 minutes to reach the same temperature. Urinary iodine levels determined by the rapid cooling method in the three samples were (158.3 ± 1.9), (133.7 ± 2.7), and (219.2 ± 3.1) μg/L, respectively, while those determined by the natural cooling method were (155.5 ± 2.7), (136.2 ± 2.3), and (215.1 ± 3.9) μg/L, respectively. There was no statistically significant difference between the results of the two methods ( t = 1.43, P = 0.192). In the validation experiments for the rapid cooling method, the linear equations for the low-concentration (0 - 300 μg/L) and high-concentration (300 - 1 200 μg/L) iodine standard series were y = - 0.003 0 x + 0.117 5 ( r = - 0.999 7) and y = - 0.001 5 x + 0.649 3 ( r = - 0.999 8), respectively. The precision experiments showed that the relative deviation values of the repeated tests for two different iodine concentrations (300 and 600 μg/L) were 1.99% and 1.90%, respectively. The accuracy experiments demonstrated that the repeated test results for the national urinary iodine reference materials (GBW09110e and GBW09109m) were 216.8 and 134.7 μg/L, respectively, with relative standard deviations of 1.04% and 1.35%, respectively, all within the reference ranges[(218 ± 15) and (135 ± 10) μg/L]. The detection limit was 1.20 μg/L. The spike recovery rate ranged from 92.90% to 99.10%, with an average recovery rate of 96.77%. Conclusions:The rapid cooling-arsenic-cerium catalytic spectrophotometry method not only shortens the time required for urinary iodine determination and improves the speed of measurement, but also provides accurate results, which can be applied when a large number of urine samples need to be measured in a short period of time.

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal hematologic disorder. Historically, most PNH patients mainly received supportive treatment, which led to poor quality of life and high mortality rates. The advent of downstream complement C5 inhibitors has dramatically changed the natural course of PNH. These inhibitors alleviate clinical symptoms, significantly reduce severe complications, and improve survival rates. Despite their ability to control intravascular hemolysis and enhance hemoglobin levels, approximately half of the treated patients still require blood transfusions due to extravascular hemolysis (EVH) mediated by upstream complement component C3. Upstream complement inhibitors not only control IVH but also reduce the deposition of C3 fragments, thereby solving the problem of EVH and further improving the clinical outcomes of PNH patients. However, due to the cascade of complement activation and the increased accumulation of PNH red blood cells after treatment with upstream complement inhibitors, there may be an increased risk of more severe breakthrough hemolysis events. Additionally, the potential risk of infections associated with complement inhibitors is another issue that needs to be addressed.

Humans ; Leukemia, Large Granular Lymphocytic/drug therapy* ; Aminopyridines ; Benzamides

Objective:To investigate the clinical characteristics of patients with acquired aplastic anemia (AA) accompanied by abnormal antinuclear antibody (ANA) and autoantibodies and their effects on the efficacy of immunosuppressive therapy (IST).Method:A retrospective case-control study was conducted, analyzing the clinical data of 291 patients with AA who underwent IST and were screened for autoantibodies at initial diagnosis between January 2018 and December 2019 at Blood Diseases Hospital, Chinese Academy of Medical Sciences. According to the titer of ANA at the initial diagnosis, extracted nuclear antigen antibodies (ENAs) abnormality and the change of ANA titer after treatment, the treatment responses of 3 months and 6 months after IST were compared. The correlation between clinical features and ANA abnormality was analyzed by univariate and multivariate logistic regression analysis. The parameters of univariate analysis P<0.1 were included in multivariate analysis, stepwise regression analysis and subgroup analysis. Results:A total of 291 patients were included in the study, of which 145 (49.83%) were male. Among all patients, 147 (50.52%) tested positive for ANA at initial diagnosis, with titers of 1∶100, 1∶320, and 1∶1 000 observed in 94, 47, and 6 cases, respectively. Female gender, older age, presence of paroxysmal nocturnal hemoglobinuria (PNH) clone, and higher levels of IgG, IgA, and thyroid hormone were significantly associated with ANA positivity at initial diagnosis, while white cell counts, reticulocytes, and free triiodothyronine were significantly lower than that of ANA-negatively patients (all P<0.05). Furthermore, logistic regression analyses revealed that female gender ( OR=1.980, 95% CI 1.206-3.277), older age ( OR=1.017, 95% CI 1.003-1.032), and presence of PNH clone ( OR=1.875, 95% CI 1.049-3.408) were independent risk factors for ANA positivity at initial diagnosis. Subgroup analysis indicated that the risk of ANA positivity at initial diagnosis was even higher in PNH clone-positive patients in the subgroups of females ( OR=1.24, 95% CI 1.02-1.51), severe AA ( OR=1.26, 95% CI 1.07-1.47), and age≥40 years ( OR=1.26, 95% CI 1.05-1.52) (all P<0.05). However, ANA titers at initial diagnosis, presence of other abnormal ENAs, and changes in ANA titers after treatment with IST were not correlated with treatment response (all P>0.05). Conclusions:Approximately 50% of patients with AA had abnormal ANA, and their presence was significantly associated with female gender, older age, and presence of PNH clone at initial diagnosis. However, the presence of abnormal ANA and changes in ANA titers after treatment did not affect the efficacy of IST in patients with AA.

Objective:To investigate the effect of on-demand glucocorticoid strategy on the occurrence and outcome of porcine anti-lymphocyte globulin (p-ALG) -associated serum sickness in aplastic anemia (AA) .Methods:The data of AA patients who received in the Anemia Diagnosis and Treatment Center of Haematology Hospital, CAMS & PUMC from January 2019 to January 2022 were collected. Among them, 35 patients were enrolled in the on-demand group, with the glucocorticoid strategy adjusted based on the occurrence and severity of serum sickness; 105 patients were recruited in the usual group by matching the age and disease diagnosis according to 1∶3 ratio in patients who received a conventional glucocorticoid strategy in the same period. The incidences, clinical manifestations, treatment outcomes of serum sickness, and glucocorticoid dosage between the two groups were analyzed.Results:The incidences of serum sickness in the on-demand group and the usual group were 65.7% and 54.3% ( P=0.237) , respectively. The median onset of serum sickness was the same [12 (9, 13) d vs the 12 (10, 13) d, P=0.552], and clinical symptoms and signs, primarily joint, and/or muscle pain, fever, and rash were similar. Severity grades were both dominated by Grades 1-2 (62.8% vs 51.4%) , with only a few Grade 3 (2.9% vs 2.9%) , and no Grades 4-5. No significant difference in the serum sickness distribution ( P=0.530) . The median duration of serum sickness was the same [5 (3, 7) d vs 5 (3, 6) d, P=0.529], and all patients were completely cured after glucocorticoid therapy. In patients without serum sickness, the average dosage of prophylactic glucocorticoid per patient in the usual group was (469.48 ±193.57) mg (0 in the on-demand group) . When compared to the usual group, the average therapeutic glucocorticoid dosage per patient in the on-demand group was significantly lower [ (125.91±77.70) mg vs (653.90±285.56) mg, P<0.001]. Conclusions:In comparison to the usual glucocorticoid strategy, the on-demand treatment strategy could significantly reduce glucocorticoid dosage without increasing the incidence of serum sickness; in addition, the duration of serum sickness and the incidence of above Grade 2-serum sickness were similar.

Objective:To analyze the diagnostic value of cell-free plasma metagenomic next-generation sequencing (mNGS) pathogen identification for severe aplastic anemia (SAA) bloodstream infection.Methods:From February 2021 to February 2022, mNGS and conventional detection methods (blood culture, etc.) were used to detect 33 samples from 29 consecutive AA patients admitted to the Anemia Diagnosis and Treatment Center of the Hematology Hospital of the Chinese Academy of Medical Sciences to assess the diagnostic consistency of mNGS and conventional detection, as well as the impact on clinical treatment benefits and clinical accuracy.Results:①Among the 33 samples evaluated by mNGS and conventional detection methods, 25 cases (75.76%) carried potential pathogenic microorganisms. A total of 72 pathogenic microorganisms were identified from all cases, of which 65 (90.28%) were detected only by mNGS. ②All 33 cases were evaluated for diagnostic consistency, of which 2 cases (6.06%) were Composite, 18 cases (54.55%) were mNGS only, 2 cases (6.06%) were Conventional method only, 1 case (3.03%) was both common compliances (mNGS/Conventional testing) , and 10 cases (30.3%) were completely non-conforming (None) . ③All 33 cases were evaluated for clinical treatment benefit. Among them, 8 cases (24.24%) received Initiation of targeted treatment, 1 case (3.03%) received Treatment de-escalation, 13 cases (39.39%) received Confirmation, and the remaining 11 cases (33.33%) received No clinical benefit. ④ The sensitivity of 80.77%, specificity of 70.00%, positive predictive value of 63.64%, negative predictive value of 84.85%, positive likelihood ratio of 2.692, and negative likelihood ratio of 0.275 distinguished mNGS from conventional detection methods (21/12 vs 5/28, P<0.001) . Conclusion:mNGS can not only contribute to accurately diagnosing bloodstream infection in patients with aplastic anemia, but can also help to guide accurate anti-infection treatment, and the clinical accuracy is high.

Objective:Short-term efficacy and safety of afatrombopag conversion therapy in patients with aplastic anemia (AA) who were previously ineffectively treated with intense immunosuppressive therapy (IST) combined with TPO receptor Agonist (TPO-RA) or who were unable to tolerate the side effects of TPO-RA.Methods:Analysis of patients with severe aplastic anemia (SAA) treated in Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College from January 2021 to December 2021 who received IST combined with TPO-RA (eltrombopag/hatrombopag) for at least 6 months, but was ineffective for at least 3 months or patients who cannot continue to use TPO-RA due to side effects, and switched from TPO-RA to avatrombopag (APAG) , and treated for at least 6 months. This study analyzed its short-term efficacy and evaluated its safety.Results:The median age was 54 (14-68) years old among the 16 patients with AA (8 male and eight female) . A total of ten patients (refractory group) who did not respond to IST combined with TPO-RA were converted to APAG median therapy for 6 (6-10) months. Only seven patients (70% ) obtained trilineage HR [four cases of complete treatment response (CTR) , one case of good treatment response (GPR) , and two cases of partial treatment response (PR) ], all of which began to take effect at 3 months of APAG treatment. There were six patients with TPO-RA intolerance, and APAG was treated for 6 (2 to 8) months. About four patients (67% ) received HR (three GPR cases and one PR case) , of which two patients received PR at 3 months and four patients received HR at 6 months of APAG treatment. No APAG-related grade 2 or more adverse events occurred during the APAG therapy, no thrombotic events occurred, no fibrologic tissue hyperplasia was found in the bone marrow pathology reexamination at 6 months of treatment, and none of the patients discontinued the drug due to adverse events.Conclusion:APAG may be a better switching treatment option for patients with AA who are refractory or are intolerant to TPO-RA.

Objective:To investigate the expression of iron-regulating erythroid factors in different types of erythropoiesis disorders.Methods:From January 2016 to November 2019, the plasma concentrations of iron-regulating erythroid factors were measured by ELISA methods in 47 patients with different types of erythropoiesis disorders. The adaptation orientation of iron-regulating erythroid factor expression with bone marrow erythropoiesis activities (represented by bone marrow-nucleated erythrocytes ratio) was analyzed.Results:The median plasma growth differentiation factor (GDF) 15 levels in patients with polycythemia vera (PV) , pure red cell aplasia (PRCA) , autoimmune hemolytic anemia (AIHA) , and myelodysplastic syndrome (MDS) were 266.01 ng/L (112.40, 452.37) , 110.63 ng/L (81.41, 220.42) , 52.11 ng/L (32.61, 171.66) , and 276.53 (132.16, 525.70) ng/L, respectively, which were significantly higher than those in normal patients with 37.45 (19.65, 57.72) ng/L (all P < 0.01) . The plasma TWSG1 expression levels were not significantly different in patients with PV, PRCA, AIHA, and MDS from those of normal patients (P>0.05) . The median plasma GDF11 level in PV was 74.75 (10.95, 121.32) ng/L, which was significantly higher than 36.90 (3.38, 98.34) ng/L in normal control subjects ( P<0.01) . However, no statistical differences were observed in the other three subjects ( P>0.05) . The median plasma erythroferrone (ERFE) levels in AIHA and PV were 121.76 ng/L (68.12, 343.11) and 129.63 (47.02, 170.03) ng/L, respectively, with the highest level in AIHA in all the studied types of erythropoiesis disorders. The bone marrow-nucleated erythrocytes ratio was significantly and positively correlated with ERFE ( r=0.458, P=0.001) but not with GDF15 ( r=-0.163, P=0.274) , GDF11 ( r=0.120, P=0.421) , and TWSG1 ( r=-0.166, P=0.269) . Conclusion:The expression profile of iron-regulating erythroid factors is not exactly the same in different types of erythropoiesis disorders. ERFE demonstrated the highest correlation with erythropoiesis activities.