Objective:To evaluate the efficacy and safety of chimeric antigen receptor T-cell (CAR-T) therapy followed by allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with Ph-like acute lymphoblastic leukemia (Ph-ALL) .Methods:Patients with Ph-ALL who underwent CAR-T therapy followed by allo-HSCT from March 2018 to August 2023 at the First Affiliated Hospital of Soochow University were included, and their clinical data were retrospectively analyzed.Results:Of the 21 patients, 14 were male and 7 were female. The median age at the time of CAR-T therapy was 22 (6-50) years. Seven patients had ABL1-like rearrangements, and 14 had JAK-STAT rearrangements. Prior to CAR-T therapy, 12 patients experienced hematologic relapse; 7 were multiparameter flow cytometry minimal residual disease (MFC-MRD) -positive and 2 were MFC-MRD-negative. CAR-T cells were derived from patients’ autologous lymphocytes. Nine patients were treated with CD19 CAR-T cells, and 12 were treated with CD19/CD22 CAR-T cells. After assessment on day 28 after CAR-T therapy, 95.2% of the patients achieved complete remission, with an MRD-negative remission rate of 75%. Nineteen patients developed grade 0–2 cytokine release syndrome (CRS) and 2 patients suffered grade 3 CRS, all cases of which resolved after treatment. All patients underwent allo-HSCT after CAR-T therapy. The median time from CAR-T therapy to allo-HSCT was 63 (38-114) days. Five patients experienced relapse after CAR-T therapy, including four with hematologic relapse and one with molecular relapse. The 3-year overall survival (OS) rates in the ABL1 and JAK-STAT groups were (83.3±15.2) % and (66.6±17.2) %, respectively ( P=0.68) . The 3-year relapse-free survival (RFS) rates were (50.0±20.4) % and (55.6±15.4) % in the ABL1 and JAK-STAT groups, respectively. There was no significant difference in 3-year OS or RFS between the two groups. Conclusions:CAR-T therapy followed by allo-HSCT leads to rapid remission in most patients with Ph-ALL and prolongs leukemia-free survival.

Objective To evaluate the changes in motor function impairment and brain functional networks of patients with acute ischemic stroke(AIS) through parameters of spontaneous activities of both upper limbs and electroencephalogram graph theory analysis methods. Methods The data of 34 acute ischemic stroke patients(observation group) with upper limb motor disorders who were treated in the Department of Neurology of the Affiliated Hospital of Southwest Medical University from January 2022 to October 2023, and 40 healthy controls (HC group) were collected. The subjects completed the National Institutes of Health Stroke Scale (NIHSS) and Fugl-Meyer Assessment (FMA) within 7 days, and wore wrist activity recorders (Actiwatch) continuously for 24 hours to collect data on spontaneous activities of upper limbs and analyzed related parameters such as the coordination coefficient of both upper limbs (r), the activity ratio of the affected side to the healthy side upper limb (ULAR), etc. At the same time, all subjects completed approximately 2 hours of 19-channel electroencephalogram examination. After preprocessing the electroencephalogram data, 5 segments of 10-second resting-state electroencephalogram were extracted for graph theory. Results ① Compared to healthy individuals, AIS patients exhibited decreased functional connectivity edges in the δ and θ bands, with substantial reductions in network connections in the α band. In the β band, connections between the frontal, right parietal, and occipital regions weakened, while connections from the right temporal lobe to the left temporal lobe strengthened. In the γ band, there was a significant increase in connections throughout the brain. ② Graph theory analysis revealed significantly increased shortest path lengths (α band: t=2.228, P < 0.05, d=-0.52; β band: t=-3.641, P < 0.01, d=-0.878) and decreased global efficiency (α band: t=2.535, P < 0.05, d=0.591; β band: t=3.321, P < 0.01, d=0.803) in the observation group compared to the control group. In the γ band, local efficiency (t=3.279, P < 0.01, d=0.765) and clustering coefficients were significantly higher (t=3.358, P < 0.01, d=0.783). ③ In the γ band, the ULAR≤30% group showed significantly reduced shortest path length (t=-2.063, P < 0.05, d=-0.802) and increased global efficiency (t=2.226, P < 0.05, d=0.865), local efficiency (t=2.95, P < 0.05, d=1.147), and clustering coefficient (t=2.962, P < 0.05, d=1.148). ④ In the observation group, the bilateral upper limb coordination coefficient during sleep was negatively correlated with NIHSS scores (r=-0.389, P < 0.05) and ULAR (r=-0.395, P < 0.05), while FMA scores were positively correlated with ULAR (r=0.442, P < 0.05). Conclusion The parameters of spontaneous activities of the upper limbs can be used to determine the impairment of motor function in AIS patients. The combination of changes in brain functional networks and motor impairments can provide new ideas for the study of their neural network mechanisms.

Animals ; Glycyrrhiza/adverse effects* ; Plant Extracts/toxicity* ; Rats ; Rats, Sprague-Dawley ; Sargassum

Objective: To investigate the expressions of miR-144 and lncRNA DNAJC3-AS1 in breast cancer tissues and their effects on chemo-resistance of breast cancer MCF-7 cells. Methods: A total of 196 pairs of breast cancer tissues and corresponding adjacent normal tissues collected between January, 2012 and December, 2016 in Department of Oncology, 3201 Hospital were used for this study. The relative expressions of DNAJC3-AS1, DNAJC3 and miR-144 in collected tissues were determined using qPCR, and their impact on the survival of BC patients was also analyzed. The targeted binding relationship between DNAJC3-AS1 and miR-144 was verified by Luciferase reporter gene assay. DNAJC3-AS1 over-expression plasmid and miR-144 mimics were transfected into MCF-7 cell lines respectively, and qPCR was used to verify the transfection efficiency. The effects of DNAJC3-AS1 and miR-144 overexpression on proliferation and cisplatin sensitivity of MCF-7 cells were verified by CCK-8 assay. Results: DNAJC3-AS1 and its host gene DNAJC3 were highly expressed in BC tissues (all P<0.01), and these two were positively correlated (r=0.451, P<0.01); in addition, patients with high expressions of DNAJC3-AS1 and DNAJC3 had a shorter survival period (all P<0.01). miR-144 was highly expressed in BC tissues (P<0.01) and negatively correlated with DNAJC3-AS1 (r=-0.524, P<0.01). The average over-expressionfold for DNAJC3-AS1 was 13.47 (P<0.01), while the fold for miR-144 was 20.27 (P<0.01). Bioinformatics analysis and fluorescence reporter gene assay confirmed that DNAJC3-AS1 could specifically bind to miR-144. MCF-7 cell lines over-expressing DNAJC3-AS1 and miR-14 were successfully constructed; compared with control group, cells in DNAJC3-AS1 over-expression group exhibited significantly enhanced proliferation and reduced cisplatin-sensitivity (all P<0.01), while the cells in miR-144 over-expression group showed significantly enhanced drug sensitivity (P<0.01). Conclusion: miR-144 and lncDNAJC3-AS1 were highly expressed in BC tissues, miR-144 promotes cisplatin sensitivity of BC MCF-7 cells through targeting DNAJC3-AS1.

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.

Objective: Early T-cell precursor acute lymphoblastic leukemia (ETP-ALL) is a recently recognized high-risk T lymphoblastic leukemia subgroup. The optimal therapeutic approaches to adult patients with ETP-ALL are poorly characterized. In this study, we explore the efficacy and outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for ETP-ALL. Methods: The clinical data of 23 patients with ETP-ALL receiving allo-HSCT from 2010 to 2018 were retrospectively analyzed. Patients with ETP-ALL were diagnosed based on the characteristic immunophenotypes. Second-generation sequencing was done in all patients. As to the donors, 12 patients had haploidentical donors (Haplo-HSCT) , 7 HLA-matched sibling donors (Sib-HSCT) and 4 HLA-matched unrelated donors (URD-HSCT) . Before transplantation, 19 patients achieved complete remission (CR) and 4 patients without. Results: The main clinical features of ETP-ALL included high white blood cell counts in 5 patients, splenomegaly in 14, lymphadenopathy in 19, and thymus masses in 5. According to cytogenetic and molecular characteristics, 11 patients had gene mutations related to myeloid tumors, and 7 with high risk Karyotype. After first induction regimen, 14/23 patients achieved CR. 5 patients reached CR after more than 2 cycles of chemotherapy, while another 4 patients did not reach CR. After allo-HSCT, 22 patients were successfully implanted. The median time of granulocyte and platelet reconstitution was +12 and +19 days. One patient died of transplant-related infection at +14 days. The estimated 18-month overall survival (OS) and relapse-free survival (RFS) rates were (55.0±14.4) % and (48.1±14.7) % respectively. Transplant-related mortality was 4.3%. The median OS in patients achieving CR before transplantation was 20 months, however, that in patients without CR was only 13 months. OS and RFS between haplo-HSCT and sib-HSCT were comparable (P=0.460 and 0.420 respectively) . Conclusions Allo-HSCT is an effective therapy in some patients with ETP-ALL. Salvage HSCT cannot overcome the poor outcome. Haplo-HSCT and sib-HSCT in ETP-ALL patients have the similar clinical outcome.

Objective: To evaluate the prognostic significance of minimal residual disease (MRD) monitoring by 10-color flow cytometry in multiple myeloma (MM) patients after treatment. Methods: 150 patients with MM who were admitted to the First Affiliated Hospital of Soochow University from July 2015 to July 2017 were retrospectively analyzed. Clinical data, MRD data monitoring by 10-color flow cytometry and prognosis were analyzed. Results: 39.1% (34/87) patients were MRD negative after induction chemotherapy, and 49.3% (34/69) patients were MRD negative within 1 year after autologous hematopoietic stem cell transplantation (ASCT) . MRD-negative patients after induction chemotherapy or after transplantation have better progress-free survival (PFS) than MRD-positive patients (P=0.022 and P<0.001) . According to the changes of MRD pre-ASCT and after ASCT, the patients were divided into 4 groups: patients with MRD continued negativity,improved from MRD positive to MRD negative, MRD continued positivity, transformed from MRD negative to MRD positive. The two-year PFS of the four groups were 83%, 82%, 44%, 0, respectively, (P=0.002) . Multivariate analysis showed that the level of MRD after induction chemotherapy was an independent factor for PFS (P=0.002) , HR=4.808 (95%CI 1.818-12.718) . Conclusion Patients with MRD negative after treatment is a better prognosis marker than complete remission or even the best marker, which can evaluate prognosis by combining R-ISS and cytogenetic changes.

Objective To establish a clinical scoring system for the prognosis of patients with single large hepatocellular carcinoma (HCC) after hepatectomy.Methods 268 patients with single large HCC who underwent hepatectomy in West China Hospital of Sichuan University from January 2009 to December 2013 were included in this prospective study. There were 227 males and 41 females, of which 198 cases aged≤60 years old, 70 aged>60 years old. The informed consents of all patients were obtained and the local ethical committee approval was received. The patients' survival were observed. The independent risk factors for postoperative prognosis of patients with single large HCC were selected by Cox proportional risk regression mode. Based on the risk factors, the prognostic scoring system for single large HCC wasestablished. The scoring system was tested through survival analysis by Kaplan-Meier and Log-rank test. Results The median overall postoperative survival was 45 months, and the tumor-free survival was 31 months. Platelet-to-lymphocyte ratio (PLR)≥107, tumor diameter≥6.8 cm and positive microvascular invasion (MVI) were the independent risk factors for postoperative overall survival and tumor-free survival in patients with single large HCC (HR=1.004, 1.092, 2.233 and 1.003, 1.062, 1.534; P<0.05). Every independent risk factor was assigned 1 point. All patients were divided into low risk group (0 point), moderate risk group (1-2 points) and high risk group (3 points). The 5-year survival rate of high risk group was 25.4％, and that of moderate and low risk group was 33.2％ and 52.1％ respectively, where significant difference was observed (χ2=23.1, P<0.05). Similar Results were observed when the scoring system was used in patients with or without cirrhosis.Conclusions PLR≥107, tumor diameter≥6.8 cm and positive MVI are the independent risk factors for the prognosis of patients with single large HCC after resection. The prognostic scoring system established in this study can be used to predict the postoperative long-term survival of patients.

Objective The aim of this study is to analyze the long-time outcome of hepatocellular carcinoma(HCC)patients with micro-vascular invasion underwent liver resection combined with transarterial chemoembolization(TACE).Methods Our database of surgical re-section from January 2009 to September 2015 was retrospectively analyzed.This study was conducted on 296 HCC patients with MVI.Patients were divided into two groups:one group underwent liver resection (n =159)and another for liver resection combined with TACE (n =137). The 5-year overall survival rate (OS)and disease free survival (DFR)were compared.A multivariate Cox proportional hazards regression a-nalysis was performed to assess the prognostic risk factors associated with overall survival rate.Results The 5-year OS and 5-year DFR see significant difference (OS:18% vs.8%,P =0.001;TRF:15% vs.8%,P =0.008).Multivariate analysis revealed that HBsAg(HR 1.596, P =0.002,95% CI 1.194 ~2.131),tumor size >5 cm(HR 0.729,P =0.042,95% CI 0.539 ~0.989)as well as multiple tumors(HR 1.480,P =0.049,95% CI 1.002 ~2.186)were correlated to poor overall survival rate.Conclusion Surgical resection combined TACE for HCC patients with MVI realized a better prognosis than patients merely underwent therapy of resection.

OBJECTIVETo explore the clinical and laboratory characteristics in favor of the diagnosis of Ph/BCR-ABL positive acute myeloid leukemia (Ph/BCR-ABL⁺ AML).

METHODSRetrospectively analyzed the clinical and laboratory characteristics of 12 Ph/BCR-ABL⁺ AML cases from Feb, 2006 to Dec, 2013, with classic myeloid blast crisis of chronic myeloid leukemia (CML-MBC) as control, and followed-up the survival in these two cohorts of patients.

RESULTSThe median age of 12 Ph/BCR-ABL⁺ AML was 27.5 years, 10 cases (83.3%) showed non/mild splenomegaly, and mainly comprised of M₂ and M₄ subtypes according to FAB classification. The median number of basophils and megakaryocytes in peripheral blood and bone marrow was lower than that of CML-CBC patients. All the cases expressed myeloid antigens, 8 cases (66.7%) expressed CD34, 11 cases were detected with t(9;22), 5 cases (45.5%) with additional chromosomal abnormalities, including 1 case of inv(16). All the cases had BCR-ABL transcripts at diagnosis:3(25.0%) cases were e1a2 type and the remaining was b2a2/b3a2type, among which 1 case coexpressed CBFβ-MYH11. Two out of 6 cases existed AML-like mutations:1 case of CEBPA and the other of FLT3-TKD. For all the patients, 7 cases achieved complete remission (CR), including 6 out of 7 cases receiving induction chemotherapy combined with tyrosine kinase inhibitor (TKI) achieved CR, and 1 out of 3 cases receiving chemotherapy alone achieved CR. The median overall survival was 16.5 months, that of allo-HSCT group was 33.5 months, which was higher than that of non-HSCT group (5.5 months).

CONCLUSIONThe expression of e1a2 type BCR-ABL, the coexpression of fusion genes which were more common in AML, the existence of AML-like mutations were all indications of a de novo Ph/BCR-ABL⁺ AML. Low induction CR rate and short survival of Ph/BCR-ABL⁺ AML implied that chemotherapy combined with TKI and followed by allo-HSCT in CR was the only effective way to improve their survival.