Donor-specific HLA antibodies (DSA) are the main cause of graft failure in patients receiving haploidentical hematopoietic stem cell transplantation (Haplo-HSCT) and mismatched unrelated-donor hematopoietic stem cell transplantation（MMUD-HSCT）. DSA leads to graft failure by destroying donor hematopoietic stem cells mainly through the action of the ADCC effect. To study the effectiveness of a desensitization regimen including low-dose total body radiotherapy (TBI) combined with intravenous immunoglobulin (IVIg) and dexamethasone, a retrospective nested case-control study was conducted in four transplant centers from September 2020 to December 2023.36 patients with DSA mean fluorescence intensity (MFI ) ≥ 5000 were used as the study subjects for the study group. A control group of 72 patients with MFI < 500 was selected and matched 2:1 to the study group for age, sex, diagnosis, disease status, donor sources, HLA-matched grafts, ABO compatibility, mononuclear cells, and CD34+ cells. All of the study groups received a desensitization program; the control group was untreated. In the study group, 12 patients with MFI ≥10,000 as a strong DSA-positive group; and 14 patients with 5000 ≤ MFI <10,000 as a DSA-positive group. The median fluorescence intensity values of HLA class I antibodies in the study group before desensitization and +14d were 11,167.94 (5,091-21,600) vs. 4,680 (0-17,764) (p<0.001), and the median fluorescence intensity values of HLA class II antibodies before desensitization and +14d were 11,012 ( 7,084-23,781) vs. 5,000 (1,100-20,370) (p<0.001).In the study group, 97.2% of patients achieved hematopoietic reconstitution, and median implantation times for neutrophils and platelets were 12 (10-28) and 13 (10-24) days, respectively. In the study group and control group patients ,+14d neutrophil and platelet cumulative implantation rates were 94.4% ± 3.8% vs 87.5% ± 3.9% (p= 0.882) and 68.6% ± 7.8% vs 69.4% ± 5.4% (p= 0.292), respectively; The incidence of graft malfunction was 8.3% vs 9.7% (p=1.000), respectively. The 2-year OS, DFS, and CIR were comparable in the study and control groups, 76.6% ± 7.6 vs 88.9% ± 4.0% (p=0.467), 76.44% ± 8.7% vs 87.7% ± 4.1% (p=0.501), and 8.2% ± 5.9% vs 7.4% ± 3.2% (p=0.899), respectively. There was a trend of higher NRM in the study group than the control group (16.5% ± 7.8% vs 9.5% ± 3.7% (p=0.324)), but the difference was not statistically significant. The difference in the incidence of aGVHD, cGVHD, and viral infections between the two groups was not statistically significant. In the subgroup analysis, there were no statistically significant differences in the incidence of 2-year OS, DFS, CIR, NRM, aGVHD, cGVHD, and viral infection in the strongly DSA-positive, DSA-positive, and control groups. Low-dose TBI combined with IVIg and dexamethasone can be effectively used to desensitize DSA-positive patients in the PT/CY transplantation mode and to overcome the effect of DSA on implantation failure of HLA-incompatible hematopoietic stem cell transplants.

目的 探讨慢性粒细胞白血病(CML)伴发非ABL1激酶区(KD)突变与小分子酪氨酸激酶抑制剂(TKI)疗效及疾病进展的关系.方法 采用二代测序技术(NGS)检测2017年3月至2021年4月于河南省肿瘤医院血液科就诊的120例CML患者108种血液肿瘤相关基因,分析非ABL1 KD突变谱及其与患者临床资料和ABL1 KD突变的关系.结果 58例(48％)CML患者检出非ABL1KD突变,其中附加染色体异常患者的突变比例(63％)显著高于无附加染色体异常的患者(34％),差异有统计学意义(P=0.003).不同分期CML患者突变比例分析中,急变期(73％)突变比例高于加速期(67％)和慢性期(42％)(P=0.031);WT1突变和转录调节类相关突变在AP期CML患者的比例(WT1+:17％,P=0.027;转录调节类突变:42％,P=0.001)均高于CP期;患者TKI治疗反应分析中,最佳反应组突变比例(30％)低于警告组(52％)和耐药组(61％)(P=0.018);RUNX1突变在TKI治疗失败组(16％)的比例高于警告组(2％)(P=0.024);非ABL1 KD突变的CML患者出现ABL1 KD突变的比例(60％)高于无伴发非ABL1 KD突变患者(36％)(P=0.022);差异有统计学意义.结论 非ABL1 KD突变与TKI疗效及疾病进展密切相关,WT1和RUNX1突变可作为CML精准诊断的分子靶点.

Introduction FLT3 internal tandem duplication (ITD) is commonly detected in patients with acute myeloid leukemia (AML) and is known to confer poor prognosis. It has different insertion numbers, mutation burden, insertion sizes, insertion sites and insertion nucleotide sequences, showing complex diversity. However, the guideline of AML did not sufficiently consider the impact of the diversity of FLT3-ITD on treatment responses, and previous studies have not been consistent with the impact of FLT3-ITD diversity on clinical prognosis. In this study we examined the diversity of FLT3-ITD and correlated these parameters with clinical outcome in adult AML. In addition, we revealed the correlation between FLT3-ITD and drug resistance through evolutionary trajectories in primary resistant patients. Methods We included adult patients from 2016-2021 at Henan Cancer Hospital with newly diagnosed adult AML. next generation sequencing (NGS) was performed for all patients at diagnosis. The FLT3-ITD was also detected using conventional methods based on PCR, with a cut-off of 3% for positivity. The comparative analysis of NGS and PCR was done to validate the diversity of FLT3-ITD. Whole genome sequencing (WGS) data for multiple time points were used to analyze clonal evolution. Data were collected by chart review. The chi-squared or Fisher's exact test, Mann-Whitney U test, log-rank and Cox regression analyses were performed. Results Among 406 newly diagnosed adult AML patients, 24.4% (99/406) patients were detected with FLT3-ITD. The FLT3-ITD diversity was validated by NGS and PCR data from 90 FLT3-ITD positive patients. 61.1% (55/90) patients had single FLT3-ITD (sFLT3-ITD) (31M/24F, median age 46 at diagnosis, range 18-64), and 38.9% (35/90) patients carried multiple FLT3-ITDs (mFLT3-ITD) at the same time (23M/12F, median age 47 at diagnosis, range 19-64). A total of 161 FLT3-ITDs were detected, with a median mutation burden of 21.51% (0.71-93.21%) and a median insertion length of 36bp (9-189 bp). 80.20% of FLT3-ITD insertions were in the near-membrane region, and 45.54% of FLT3-ITD insertions contained exogenous nucleic acid sequences. Patients with mFLT3-ITD had a significantly lower CR rate than those with sFLT3-ITD (25.7% vs 60.0%, p=0.001), while the CR rate of patients with sFLT3-ITD was similar to patients without FLT3-ITD (60.0% vs 58.6%, p=0.521). In the setting of NPM1, the CR rate of patients with sFLT3-ITD+NPM1 mut improved significantly when compared to patients with sFLT3-ITD+NPM1 wild (76.9% vs 44.8%, p=0.015). Unfortunately, the state of NPM1 had no impact on the CR rate of patients with mFLT3-ITD. After a median follow-up of 38.9 months, the median overall survival (OS) has not been reached for patients with a single FLT3-ITD, and the median OS for patients with multiple FLT3-ITDs was only 12.4 months (95 CI, 3.8 to 20.9) ( p=0.028). Multivariate analysis confirmed its prognostic significance on CR rate (HR 2.13, 95% CI, 1.38-4.20, p=0.020) but not on OS. The mutation burden, insertion sizes, insertion sites and insertion nucleotide sequences of FLT3-ITD had no significant impact on the CR rate or OS. To explore the clonal architecture of mFLT3-ITD, single cell DNA sequencing on one patient showed that respective FLT3-ITD variants were detected in completely separated cell populations. This suggested that mFLT3-ITD independently occurred in different subclone. WGS data of persistent primary resistant patients at multiple time points were used to analyze the evolutionary trajectory under chemotherapy. We observed complete separation of evolving trajectory of different FLT3-ITD variants over the course of chemo-treatment. Importantly, FLT3-ITD subclone of diagnosis evolved into the dominant clone at resistance. Conclusion In summary, by exploring the impact of the FLT3-ITD diversity on clinical prognosis and the evolutionary trajectory of mFLT3-ITD, the FLT3-ITD number may be a biomarker to refine the risk stratification and further identify adverse populations among FLT3-ITD positive AML patients.

Background. Multiple myeloma (MM) presently remains largely incurable. The impor-tance of circular RNAs (circRNAs) in various malignancies, including MM, has been demonstrated for decades. Our goal is to decipher the complex molecular mechanism of circ_0111738 in modulating MM progression.Methods. Circ_0111738 and miR-1233-3p expressions in the collected MM cells and bone marrow aspirates were examined by qRT-PCR. CCK-8, transwell migration and invasion, and tube formation assays were performed to evaluate MM cell proliferation, migration and invasion, and angiogenesis, respectively. A tumor xenograft experiment was performed to validate the biofunction of circ_0111738's in vivo . The predicted interaction of circ_0111738 and miR-1233-3p's was determined by RNA immunopre-cipitation (RIP) and luciferase reporter assays. Apoptosis-associated proteins and the HIF-1 pathway were investigated by western blotting. Results. Circ_0111738 was poorly expressed in MM cells and patients. Overexpression of circ_0111738 reduced MM cell proliferation, migration, invasion, and angiogenesis while circ_0111738 led to opposite results. The anti-tumorigenic effect of circ_0111738 overexpression was also observed in vivo . RIP and luciferase experiments demonstrated that circ_0111738 interacted with miR-1233-3p in MM cells. The silencing of miR-1233-3p prevented the stimulation of malignant behaviors of MM cells induced by circ_0111738 silencing, including the expression of HIF-1 & alpha;.Conclusion. Our data suggest that circ_0111738 functioned as a competing endoge-nous RNA (ceRNA) and suppressed the oncogenic function of miR-1233-3p in MM by inactivating the HIF-1 pathway. Therefore, up-regulation of circ_0111738 may be a promising therapy against MM.& COPY; 2023 Instituto Mexicano del Seguro Social (IMSS). Published by Elsevier Inc. All rights reserved.

Objective To determine molecular basis of a rare HLA-A typing results carrying triple A alleles in potential allo-HSCT donor and her family. Methods HLA-A, -B, -C, -DRB1, -DQB1, -E, -F, -G of 5 members in the family were genotyped at a high-resolution level using next-generation sequencing (NGS). HLA-A of probosita was re-checked using polymerase chain reaction-sequence-based typing (PCR-SBT), and SNP oligonucleotide probes (SNP-array)were scanned with genomic DNA of probosita. Results There was 162.9Kb duplication in 6p22.1(29, 803, 377-29, 966, 301)of probosita who carried triple A alleles A*02∶01∶01, A*11∶01∶01, A*24∶02∶01. Other two family members were found to carry this haplotype: A*02∶01∶01, A*24∶02∶01, B*54∶01∶01, C*01∶02∶01, DRB1*04∶05∶01, DQB1*04∶01∶01, E*01∶01∶01∶03, F*01∶01∶01, G*01∶01∶01∶01, which as a Mendelian gene was segregated and stably transmitted through two generations. Conclusion Tiny gene duplication induces one haplotype carries two HLA-A alleles in a potential healthy donor for allo-transplantaion and stably transmits through two generations.Routine HLA typing laboratories should pay more attention to this situation and accurately report.

We sought to extend our observation of LncRNA ADAMTS9-AS1 and to specifically uncover its role on the stemness of lung adenocarcinoma (LUAD) cancer cells. ADAMTS9-AS1 was poorly expressed in LUAD. The high ADAMTS9-AS1 expression was positively associated with overall survival. ADAMTS9-AS1 overexpression attenuated the colony-forming capacity and reduced stem cell-like population of LUAD cancer stem cells (CSCs). Furthermore, ADAMTS9-AS1 overexpression increased E-cadherin expression in addition to the downregulated expressions of Fibronectin and Vimentin in LUAD spheres. In vitro results also confirmed the ADAMTS9-AS1's inhibitory effect on the growth of LUAD cells. Moreover, the antagonistic repression of miR-5009-3p levels with the expression of ADAMTS9-AS1 and NPNT was confirmed. Finally, ADAMTS9-AS1 overexpression curbed the increasing stemness of LUDA-CSC caused by NPNT silencing, thus leading to the suppression of LUAD progression in vitro. Conclusively, ADAMTS9-AS1 negatively controls the LUAD cancer cell stemness progression through regulating miR-5009-3p/NPNT axis.

Introduction The SET binding protein 1 (SETBP1) gene is an oncogene located on chromosome 18q12.3, which is related to DNA replication and gene transcription regulation. Variants in SETBP1 are involved in germline and somatic mutations, leading to extremely different pathologic consequences. Germline mutations in SETBP1 are associated with Schinzel-Giedion syndrome (SGS) and SETBP1 Haploinsufficiency Disorder. In contrast, somatic mutations in SETBP1 are linked with myeloid malignancies and clonal hematopoiesis. The vast majority of reported germline SETBP1 mutations were located in the SKI homologous region (codons 868-871). However, germline SETBP1 mutations outside the hotspot region were rarely reported and the potential carcinogenicity remained unclear. Methods Patients from 2018-2022 at the Hennan Cancer Hospital were identified. Bone marrow samples at diagnosis of 285 sporadic patients with myeloid malignancies were sequenced using next generation sequencing (NGS). The candidate SETBP1 mutations were validated by Sanger sequencing from bone marrow and exfoliated buccal epithelial cell samples. Whole exome sequencing (WES) was performed on patient 5 and her parents from the family with SETBP1 mutations. Results In our study, 8 (2.8%) patients (6 AML and 2 MDS) were identified with germ line SETBP1 mutations (Table 1, Figure 1). Importantly, patient 2 and patient 5 carried the same germ line SETBP1 p.Ser373Ala mutation. Meanwhile, 4 patients had somatic SETBP1 mutations at hotspots (p.Asp868Asn in 1 patient and p.Gly870Ser in 3 patients). None of these germ line SETBP1 mutations was detected in 597 healthy Chinese individuals except p.Ser567Phe (2 out of 597) (Table 1). Minimum allele frequencies (MAF) of the germ line SETBP1 mutations were negligible in public databases (Table 1). A significant difference ( p＜0.05) was observed in the minimum allele frequency (MAF) of germ line SETBP1 mutations between sporadic MDS/AML patients and healthy controls, which suggested that germ line SETBP1 mutations should be closely related to MDS/AML. Furthermore, we investigated the family history of the patients with germ line SETBP1 mutation. The father of patient 5 was diagnosed with MDS and carried germ line SETBP1 p.Ser373Ala mutation. Known germ line mutations or somatic mutations associated with myeloid malignancies were not detected in the family of patient 5 by WES. Patient 2, who also carried the germ line SETBP1 p.Ser373Ala mutation, had no family history of myeloid disorders. The elder sister (carrier) of patient 2 had persistent abnormal CBC while the little sister (carrier) had normal CBC. Somatic SETBP1 mutations conveyed a poor prognosis in myeloid malignancies. And in our data, patient 5 and patient 2 both appeared to be induction failure and early relapse. Then patient 2 underwent HLA-full-matched allo-HSCT in the second remission and the doner was his little sister. Unfortunately, after allo-HSCT the CR only maintained 2 months. None of the other 6 sporadic MDS/AML patients with germ line SETBP1 mutation had a family history of hematologic disorders. Many family members were detected to carry germ line SETBP1 mutations but they did not show the characteristics of myeloid disorders other than abnormal percentages of neutrophils and lymphocytes. Conclusion In our study, we identified novel germ line SETBP1 mutations in 8 sporadic MDS/AML patients, and a family history of MDS/AML was noted in one. Germ line SETBP1 mutations are not infrequent among non-familial MDS/AML patients. Identification of these cases is imperative especially in patients who are candidates for HSCT from a family donor, since the association between germ line SETBP1 mutations and recipient outcomes was uncertain.

Mucormycosis is a conditionally pathogenic fungal disease with high morbidity that mainly affects patients with decreased immunity. Diagnosis relies on the histopathological examination of microorganisms with the typical structure of mucormycetes in tissues and subsequent confirmation via culture. Early detection of causative microorganisms is critical to rapidly administer appropriately targeted antibiotics. Metagenomic next-generation sequencing (mNGS) is an innovative and sensitive technique used to identify pathogenic strains. Here we used mNGS to timely diagnose an infection with Lichtheimia ramosa and Mucor irregularis in two patients with hematologic malignancies; the infections manifested as nasal and cutaneous infections and developed after chemotherapy and small molecule targeted therapy. Following treatment with amphotericin B cholesteryl sulfate complex, the symptoms were reduced significantly, and both patients obtained successful outcomes. Additionally, we searched and summarized the current medical literature on the successful diagnosis of mucormycosis using mNGS. These cases indicated that mNGS, a novel culture-independent method, is capable of rapid, sensitive, and accurate identification of pathogens. mNGS may be a complementary method for the early identification of mucormycosis, allowing for appropriate and timely antibiotic administration and thus improving patient outcomes.

Background: The emergence of small molecule tyrosine kinase inhibitors (The Tyrosine Kinase Inhibitors, TKI) has gradually transformed chronic myelocytic leukemia (CML) from a fatal malignant disease into a chronic disease, while ABL1 kinase Kinase Domain (KD) mutation is the main reason for poor efficacy or resistance to TKIs. In recent years, with the clinical application of Next Generation Sequencing (NGS) technology, it has been found that CML patients are often accompanied by non-ABL1 kinase region gene mutations, but whether these accompanying mutations are related to the efficacy of TKIs and disease progression in CML? still uncertain. This study retrospectively analyzed the correlation between CML with non-ABL1 KD mutation and TKI efficacy and disease progression, in order to provide basis and scientific guidance for further accurate diagnosis and treatment of CML patients. Aims: To investigate the relationship between CML associated with non ABL1 Kinase Domain (KD) mutation and TKI efficacy and disease progression. Methods: 108 blood tumor related genes in 120 patients with CML were detected by Next-generation sequencing (NGS). The non ABL1 KD mutations and their relationship with relevant clinical information and ABL1 KD mutation were analyzed. Results: Non ABL1KD mutations were detected in 58 patients (48%), and the mutation proportion of patients with additional chromosome abnormalities (63%) was significantly higher than that of patients without additional chromosome abnormalities (34%) (P=0.003). In the analysis of mutation proportion in patients with different stages of CML, the mutation proportion in blastic phase(73%), accelerated phase (67%) and chronic phase (42%) decreased gradually (P=0.031). The proportion of WT1 mutation and transcriptional regulation related mutations in patients with AP CML (WT1+: 17%, P=0.027; Transcriptional regulatory mutations 42%, P=0.001) were higher than those in CP. In the analysis of TKI treatment response, the proportion of mutations detected in the best response group (30%), warning group (52%) and drug resistance group (61%) increased gradually (P=0.018). The proportion of RUNX1 mutation in TKI treatment failure group (17%) was higher than that in warning group (2%) (P=0.029). The proportion of ABL1 KD mutation in CML patients with non ABL1 KD mutation (60%) was significantly higher than that in patients without non ABL1 KD mutation (36%) (P=0.022). Summary/Conclusion: Non ABL1 KD mutations are closely related to TKI efficacy and disease progression, while WT1 and RUNX1 mutations may be new molecular targets for accurate diagnosis of CML.

Abstract Introduction Copy number aberrations (CNAs) constitute the backbone of the diagnostic classification and risk stratification in patients with acute myeloid leukemia (AML) or myelodysplastic syndromes (MDS). Although conventional cytogenetic approaches has been implemented in clinical routine to detect recurrent genomic abnormalities in AML/MDS, it presents a few obvious shortcomings, such as time-consuming in vitro cell-culturing procedure, low resolution (no smaller than 10 Mbp), and subjective bias introduced from the visual interpretation by cytogeneticists. With the technological advances of next generation sequencing (NGS), the longstanding conventional cytogenetic test is likely to be challenged by whole-genome sequencing (WGS), which is a more unbiased method to detect all types of genomic aberrations not only those well-characterized clinically actionable mutations but also cryptic alternations yet to be defined and annotated. Methods In this study a shallow WGS (sWGS) was utilized to depict genomic profiles of CNAs for 113 patients with AML/MDS, 103 of whom had undergone cytogenetic test. Bone marrow was collected and mononuclear cells were separated to harvest genomic DNA which was sequenced without capture at approximately 1x coverage depth. Genome-wide CNAs greater than 5 Mbp was analyzed for each sample. European Leukemia Net (ELN)-defining CNAs were selected and used to assign patients to a genetic risk group through the same classification systems that are used for cytogenetic test. Results sWGS identified 156 DNA segments with CNAs in 68 (60.2%) of all 113 patients. At chromosome arm level, most recurrent events were gains of chr 8 (observed in 9 AML and 6 MDS patients), losses of chr Y (observed in 4 AML and 1 MDS patients) which are consistent with previously reported cytogenetic events in AML/MDS. With 103 patients whose cytogenetic test results were available, 79 CNAs were described by cytogenetic test. Compared between cytogenetic test and sWGS, a total of 52 CNAs (65.8%) were unambiguously matched. 42 and 32 CNAs were identified by sWGS only and cytogenetic test only respectively. Of the 42 aberrations detected only by sWGS, 19 (45.2%) were shorter than normally detected by cytogenetic test (<10 Mbp). Next, the concordance between sWGS and cytogenetic test for ELN-defining CNAs [del(5q), del(17p), and monosomy 7] was specifically evaluated. A total of 22 ELN-defining CNAs events were detected, 12 (54.5%) of which were concordantly identified by both sWGS and cytogenetic test whereas the other 10 (45.5%) were solely reported by sWGS but not detected by cytogenetic test. In regard to risk-group assignments based on ELN sWGS and cytogenetic test results, sWGS provided new genetic information in 16 (15.5%) patients and six (5.8%) patients were reassigned to a different risk category due to new adverse-risk findings that were identified by sWGS. Noteworthily, concordant CNA profiles were observed between the genomic DNA of bone marrow cells and the cell-free DNA (cfDNA) of plasmas from the same patients, which indicates peripheral blood can be a less-invasive alternative to characterize CNAs by sWGS analysis so that bone marrow aspiration can be spared. Conclusions In this study, a streamlined sWGS provided an accurate, convenient and cost-effective approach to describe genomic profiles of CNAs in patients with AML/MDS. It also brought greater diagnostic yield and risk stratification information than cytogenetic test based on the standard ELN categories. Disclosures Zhu: Clinical Laboratories, Shenyou Bio: Current Employment. Zhang: Clinical Laboratories, Shenyou Bio: Current Employment. Chen: SeekIn Inc: Current Employment, Current holder of individual stocks in a privately-held company. Li: SeekIn Inc.: Current Employment, Current holder of individual stocks in a privately-held company. Chang: Clinical Laboratories, Shenyou Bio: Current Employment. Mao: SeekIn Inc: Current Employment, Current holder of individual stocks in a privately-held company.

[目的]探讨PD-L1单抗对EGFR敏感性突变肺癌细胞PD-L1的表达及对增殖的影响.[方法]选取2017年5月至2018年8月在我院呼吸内科治疗的非小细胞肺癌患者60例为研究对象,同时选取同期来我院检查的健康人群30例为对照.采集外周静脉血,测定血清标本中游离PD-L1 (sPD-L1)蛋白含量.建立肺癌细胞与T细胞共培养体系.[结果]肺鳞癌组与肺腺癌组sPD-L1的表达水平明显高于对照组(P＜0.001),而肺鳞癌组与肺腺癌组sPD-L1表达水平无统计学差异(t=2.584,P=0.086);野生组与突变组sPD-L1表达水平亦无统计学差异(t=-2.215,P=0.097).sPD-L1表达量与患者性别、年龄、分期无相关性(P＞0.05),而与患者的吸烟史、肿瘤转移相关(P＜0.05).EGFR敏感型sPD-L1表达率明显高于EGFR野生型(t=12.365,P＜0.001).加入PD-L1单抗后,EGFR的sPD-L1的表达水平有明显下降趋势,而EGFR野生型与治疗前无统计学差异(t=2.154,P=0.958).PD-L1单抗使用后,PC9、HCC827等EGFR敏感型细胞株共同体系中T细胞明显增加.[结论]sPD-1表达与EGFR敏感性无明显相关性,而与患者是否吸烟、肿瘤转移相关.与此同时,EGFR敏感突变肺癌细胞行PD-L1单抗干预后,sPD-L1的表达量明显降低,T细胞增殖明显.

Non-small cell lung cancer (NSCLC) is accounted for 80% to 85% of the total lung cancer cases and still a difficult problem to solve at present. The present study was aimed to explore the effect of S100A6 on the proliferation, invasion, migration and angiogenesis in lung cancer cell lines with the change of miR-193a expression and P53 acetylation. The expression of S100A6, CDK2, cyclinD1, VEGF, ANGII, anti-acetylp53 (K373), K-AC, P21 and Noxa were analyzed by western blot analysis. RT-qPCR analysis was used to confirm the transfection effects. CCK-8 assay and flow cytometry were reflecting the cell proliferation. Wound healing assay and transwell assay were evaluating the cell invasion and migration. The dual-luciferase reporter assay was to confirm the S100A6 as a target of miR193a. Immunofluorescence and immunohistochemical analysis were analyzing the S100A6 expression in cells and tumor tissues, respectively. As a result, S100A6 expression was increased in lung cancer cell lines and S100A6 expressed the highest in A549 cells which was chosen for the subsequent experiment. S100A6 overexpression promoted the proliferation, invasion, migration and angiogenesis of lung cancer cells with the promotion of degradation of P53 acetylation. In addition, S100A6 was demonstrated to be a target of miR193a. Moreover, miR193a expression was decreased in lung cancer cell lines and miR193a expressed the lowest in A549 cells which was chosen for the subsequent experiment. And, miR193a overexpression inhibited the proliferation, invasion, migration and angiogenesis of lung cancer cells with the enhancement of P53 acetylation. The effects of S100A6 overexpression and miR193a overexpression on tumor growth in vivo experiments were the same with that in the cell experiments. In conclusion, this study indicated that S100A6 overexpression could promote the proliferation, invasion, migration and angiogenesis of lung cancer cells by inhibiting the P53 acetylation and miR193a overexpression could reversed the above effects by decreasing the S100A6 expression in both vitro and vivo experiments.

BACKGROUND:Casitas B-lineage lymphoma proto-oncogene-b (CBLB) influences the threshold of T cell activation and controlling peripheral T cell tolerance. In the present study, we hypothesize that potentially functional single nucleotide polymorphisms (SNPs) in CBLB are associated with clinical outcomes in patients advanced non-small cell lung cancer (NSCLC) treated with the first-line chemotherapy.METHODS:We genotyped three SNPs (rs2305035, rs3772534 and rs9657904) at CBLB in 116 advanced NSCLC patients with progression free survival (PFS) data and 133 advanced NSCLC patients with overall survival (OS) data, and we assessed their associations, 95% confidence interval (CI), with clinical outcomes by using Cox proportional hazards regression analyses. In silico functional analysis was also performed for the SNPs under investigation.RESULTS:We found that associations between the three SNPs and PFS/OS were not significant in the overall NSCLC patients. The rs2305035 AA genotype was associated with a worse PFS in female patients and those of non-smokers or light smokers (95% CI, 1.14-11.81, P=0.030; 95% CI, 1.42-10.24, P=0.008; and 95% CI, 1.39-9.93, P=0.009; respectively), compared with the GG+AA genotypes. We also found that the rs9657904 CC genotype was significantly associated with a worse OS than TT + TC genotypes in male advanced NSCLC patients. Further in silico functional analysis revealed that the rs965704 T allele was significantly associated with lower mRNA expression levels of the CBLB gene.CONCLUSIONS:Our findings identified two CBLB SNPs (rs2305035 and rs9657904) that were significantly associated with PFS and OS in several subgroups of Chinese advanced NSCLC patients after the first-line chemotherapy.

Aim:To evaluate the value of detecting invasive candida infection using 1,3-β-D glucan(BG),mannan (Mn),anti-Mn IgG and IgM antibodies(Mn-IgG and Mn-lgM) in hematologic malignancies patients.Methods:Peripheral blood samples were collected from 64 patients with hematologic malignancies in the department of hematology and 45 objects in health examination from October 2015 to February 2017.Using kinetic chromogenic method and ELISA method,properties including sensitivity,specificity and so on of all the above indicators were comparative analyzed,and the impact of interference factors including neutropenia and empirical antifungal agent was evaluated in invasive candidiasis.Results:The sensitivity and specificity of BG and Mn test were 70.3％,80.0％ and 56.3％,93.3％,respectively.The continued BG test within 48 h was more specific than other indicators,but there also existed false negative result.Sensitivities of BG and Mn were both affected by neutropenia and antifungal agent.The sensitivities of Mn-IgG and Mn-lgM were lower (29.7％ and 39.1％).However,antibody detection was not affected by the above two interference factors,thus they could make up for the sensitivities of BG and Mn.AUC of the combined detection(0.830) was higher than the AUC of any single indicator.Conclusion:Combined detection of BG,Mn and anti-Mn antibodies could overcome the interference factors such as neutropenia and antifungal agent,is a more sensitive and specific test in diagnosis of invasive candidiasis.

BACKGROUND Non-small cell lung carcinoma (NSCLC) mainly includes lung squamous cell carcinoma and adenocarcinoma. This study aimed to investigate the difference between the expression of Cbl-b in lung squamous cell carcinoma and adenocarcinoma. MATERIAL AND METHODS The clinical features and survival data of NSCLC patients and Cbl-b mRNA (FPKM) were obtained from the TCGA database. Then, lung squamous cell carcinoma and adenocarcinoma cell lines were transfected with lentivirus-mediated RNA interference vector to knockdown the expression of Cbl-b. Next, a Transwell assay was performed to study the effect of Cbl-b shRNA on migration and invasion of lung squamous cell carcinoma and adenocarcinoma cells. Finally, Western blot analysis was performed to measure the expressions of PI3K, p-PI3K, AKT, p-AKT, ERK1/2, p-ERK1/2, GSK3β, p-GSK3β, mTOR, and p-mTOR protein in lung adenocarcinoma and squamous cell carcinoma cells. RESULTS The correlation of Cbl-b expression and OS was different between NSCLC adenocarcinoma and squamous carcinoma. After transfection, the expression of Cbl-b was inhibited in A549, H1975, and SW900 cells. Cbl-b shRNA promoted the migration and invasion of lung adenocarcinoma A549 and H1975 cells, but it inhibited the invasion of lung squamous cell carcinoma SW900 cells. In addition, Cbl-b regulated the expression of PI3K and ERK1/2-GSK3β pathway proteins in A549 and SW900 cells. CONCLUSIONS The OS of Cbl-b mRNA low expression in lung adenocarcinoma and squamous cell carcinoma was different. The difference in signal pathways may be one of the reasons for the difference in the correlation between Cbl-b expression and the survival rate of these 2 pathological types of lung cancer.

Breast cancer was a malignant tumor seriously threatening the life of women in the world. But the prognosis of breast cancer patients was not so satisfactory due to the limited effective therapeutics. The heterogeneity decided that more than one gene or one signaling pathway was responsible for the initiation or progression of breast cancer. DDX51 gene was a member of RNA helicases family in charge of regulation of RNA metabolism. And DDX51 gene was shown to promote proliferation in NSCLC. But we firstly reported the abundant expression of DDX51 gene in both the breast cancer tissues and cell lines in this study. And DDX51 expression was shown to be associated with TNM stage and prognosis in breast cancer patients. When DDX51 was successfully knocked down, either proliferation or DNA synthesis of MCF-7 cells was inhibited. But the ability of migration and invasion of MCF-7 cells was not affected by DDX51 gene. Furthermore, DDX51 knockdown was accompanied by inhibition of Wnt/β-catenin signaling because expression of critical members such as β-catenin, cyclin D1, TCF/LEF, and DKK1 were all affected. Therefore, this study proved that DDX51 gene promoted proliferation in MCF-7 cells by regulating Wnt/β-catenin signaling pathway and showed clinical significance in breast cancer. This study provides us a new promising hope for treatment of patients with breast cancer.

Interleukin-2 (IL-2) has been extensively used to boost the body's immune cells, especially T cells. IL-2 is a cytokine that for many years was used to activate and amplify T cells. Due to its potent T cell growth-inducing functions in vitro, for many years, IL-2 was used for the culture and expansion of various T cell products, including tumor-infiltrating lymphocytes (TIL), T cell receptors T cells (TCR T), or genetically engineered cells with chimeric antigen receptors T cells (CAR T). Despite its positive effect on T cell production, the side-effect is not well studied. Here, we reported that long-term culture with IL-2 promotes terminal differentiation and impairs rather than boosts the function of chimeric antigen receptor T cells. However, short-term culture with IL-2 predominantly generates memory CAR T cell favorable for cancer treatment.

Aim: To analyze the specific repertoire skewing and amino acid sequences of the complementarity determi-ning region 3(CDR3) of T cell receptor(TCR) β chain in metastatic lymph node from patients with breast cancer. Methods:A total of 24 TCR β chain variable region(BV) subfamilies were amplified by RT-PCR, and the CDR3 lengths were analyzed with GeneScan method in lymph nodes from 2 patients with reactive hyperplasia and metastatic lymph nodes from 5 patients with breast cancer. Sequence analysis of the CDR3 region in monoclonal or oligoclonal expanded T cells was performed. Re-sults: There were only multiclonal expanded T cells in lymph nodes from patients with reactive hyperplasia, while there were monoclonal, oligoclonal, and multiclonal expanded T cells in metastatic lymph nodes from patients with breast cancer. TCR BV presented specific repertoire skewing in metastatic lymph node from patients with breast cancer, among which only 2-5 TCR BV subfamilies were identified. The monoclonal or oligoclonal expanded T cells had different CDR3 amino acid se-quences. Conclusion: There is specific repertoire skewing of TCR BV subfamilies in metastatic lymph nodes from patients with breast cancer. The amino acid sequences of CDR3 in different T cell clone proliferation are mostly different.

目的 分析乳腺癌微转移淋巴结T细胞克隆性增生及TCR α链谱型偏移情况,了解抗肿瘤T细胞克隆的分子特征.方法 利用RT-PCR扩增10例乳腺癌微转移淋巴结T细胞32个TCR可变区(AV)亚家族的基因,基因扫描检测T细胞克隆性及TCR AV亚家族取用情况,对单克隆家族行PCR扩增TCR α链全长序列,构建重组质粒后测序,分析互补决定区(CDR3)序列.结果 乳腺癌淋巴结微转移患者T细胞呈单、寡克隆、寡克隆趋势和多克隆增生,不同患者表达1～4个TCR AV亚家族.克隆性T细胞的CDR3氨基酸序列不同,但是病例4和病例8含有相同的CDR3氨基酸基序:AM和DDKⅡ.结论 乳腺癌TCR AV基因表达具有多样性特点,TCR AV家族的取用可能与乳腺癌肿瘤抗原的多样性和不同个体的免疫应答反应有关.相同CDR3氨基酸基序的发现可能对乳腺癌的T细胞介导免疫治疗提供帮助.