Objective:To explore the characteristics and clinical significance of clonal heterogeneity in patients with acute lymphoblastic leukemia(ALL).Methods:From January 2016 to June 2019, 170 newly diagnosed ALL patients were enrolled in the Department of Hematology, Henan Cancer Hospital, including 93 males and 77 females, with a median age of 17 (2-80) years. Fifty-two ALL-related genes were detected by high-throughput sequencing technique. The clonal heterogeneity of mutations was analyzed according to the variant allele frequency (VAF) and the results of flow cytometry. The prognostic value of mutations was also evaluated.Results:Gene mutations were detected in 121 (71.2%, 121/170) patients, of which 2 or more clones were detected in 18 (52.9%, 18/34) T-cell acute lymphoblastic leukemia patients, while only 23 (16.9%, 23/136) B-cell acute lymphoblastic leukemia patients were positive of multiple mutations ( P<0.01).Gene mutation-related clonal heterogeneity analysis showed that 2 or more clones were frequent in patients with NOTCH1 mutations (13/19 patients) ( P<0.01). Event free survival (EFS) in patients with 3 or more clones was significantly lower than other patients (χ 2=10.330, P=0.016). Child ALL patients had similar result, that multiple clones predicted lower overall survival (OS) and EFS (OS: χ 2=7.974, P=0.047; EFS: χ 2=10.860, P=0.013). Conclusion:Clonal heterogeneity in ALL patients is closely related to the different origin of lymphocyte lineages and the age of onset, which may reveal the nature of the disease and predict the clinical outcome.

Objective: To explore the relationship between driver gene mutation (JAK2, MPL and CALR) and disease type in BCR-ABL negative myeloproliferative neoplasms (MPNs) including primary myeloid fibrosis (PMF), essential thrombocytosis (ET) and polycythemia vera (PV). Methods: A total of 32 MPN related genes were detected by high-throughput sequencing in 156 MPN patients. The relationships between disease type and patients′ general performance, the characteristics of driver gene mutations, concomitant gene mutations were analyzed. Results: In the population with JAK2 V617F positive mutation, the proportion of patients over 60 years old in PMF was higher than that with ET or PV. By high-throughput sequencing, 22 concomitant gene mutations were detected in 46 patients with JAK2, MPL or CALR mutations, including 4 (8.3%) in PV, 20 (29.4%) in ET, and 22 (55.0%) in PMF. DNMT3A mutation was detected only in patients with PV, while splicing factor related genes including SF3B1, SRSF2 and U2AF1 were only accompanied by PMF. According to the variation allele frequency (VAF) value of JAK2 V617F mutation, the VAF value associated with PV was the highest (68.15%), followed by PMF (37.7%) and ET (23%). However, there were significant differences in the incidence of JAK2 V617F homozygous among 3 different diseases. In patients with JAK2 mutation, the proportion of other gene mutations in PV and ET was significantly lower than that in PMF. Conclusions Under the condition of common driver gene mutations (JAK2, MPL and CALR), patients′ age, VAF value and homozygous state, concomitant gene mutations are closely related to different disease type. These correlations help to improve clinical understanding of disease characteristics and risk assessment.

ObjectiveTo investigate the efficacy and safety of camrelizumab monoclonal antibody combined with molecular-targeted therapy in elderly patients with unresectable or advanced hepatocellular carcinoma （HCC）. MethodsA retrospective analysis was performed for the patients with unresectable/advanced HCC who attended six hospitals from January 1， 2019 to March 31， 2021， and all patients received camrelizumab monoclonal antibody treatment， among whom 84.8% also received targeted therapy. According to the age of the patients， they were divided into elderly group （≥65 years） and non-elderly group （<65 years）. The two groups were assessed in terms of overall survival （OS）， progression-free survival （PFS）， objective response rate （ORR）， disease control rate （DCR）， and immune-related adverse events （irAE）. The chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups； the independent samples t-test was used for comparison of normally distributed continuous data， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. The Kaplan-Meier method was used for survival analysis， and the log-rank test was used for comparison of survival curves. Univariate and multivariate Cox proportional hazards regression analyses were used to determine the independent influencing factors for PFS and DCR at 6 months. ResultsA total of 99 HCC patients were enrolled， with 27 in the elderly group and 72 in the non-elderly group. The elderly group had an OS rate of 67.8%， an ORR of 44.4%， and a DCR of 74.1% at 12 months and a median PFS of 6.4 （95% confidence interval ［CI］： 3.0‍ ‍—‍ ‍12.4） months， with no significant differences compared with the non-elderly group （all P>0.05）. The median OS was unavailable for the elderly group， while the non-elderly group had an OS of 18.9 （95%CI： 13.0‍ ‍—‍ ‍24.8） months； there was no significant difference between the two groups （P=0.485）. The univariate and multivariate Cox regression analyses showed that major vascular invasion （MVI） was an independent risk factor for PFS （hazard ratio ［HR］=2.603， 95%CI： 1.136‍ ‍—‍ ‍5.964， P=0.024） and DCR （HR=3.963， 95%CI： 1.671‍ ‍—‍ ‍9.397， P=0.002） at 6 months， while age， sex， etiology of HBV infection， presence of extrahepatic metastasis， Child-Pugh class B， and alpha-fetoprotein>400 ng/mL were not associated with PFS or DCR at 6 months. For the elderly group， the incidence rates of any irAE and grade 3/4 irAE were 51.9% and 25.9%， respectively， with no significant differences compared with the non-elderly group （P>0.05）， and skin disease was the most common irAE in both groups （39.4%）. ConclusionCamrelizumab monoclonal antibody combined with molecular-targeted therapy has similar efficacy and safety in patients with unresectable/advanced HCC aged ≥65 years and those aged <65 years. MVI is associated with suboptimal response to immunotherapy and poor prognosis.