The value of ultrasound-guided fine needle aspiration in the diagnosis of diffuse large B-cell lymphoma: A single center experiment.

OBJECTIVES: Flow cytometry (FC) is a critical diagnostic approach for guiding targeted chemotherapy and cellular immunotherapy for relapsed and refractory lymphoma patients. The aim of the study was to investigate the value of ultrasound-guided fine needle aspiration (FNA) to improve the quality of FC specimens in relapsed and refractory diffuse large B-cell lymphoma (R/R DLBCL). METHODS: Twenty patients with R/R DLBCL after standard treatment were included. The primary lesions of all cases were confirmed by pathology. FNA and core needle biopsy (CNB) were both used for ultrasound-guided puncture, the specimens obtained by FNA are directly examined by FC, and the specimens by CNB were subjected to FC after grinding. The accuracy of FC with the two methods were evaluated using histopathology as the gold standard. RESULTS: Of the 20 R/R DLBCL cases, 19 were diagnosed as DLBCL pathologically and one was diagnosed as inflammatory granuloma. Among the specimens obtained by CNB, 14 cases examined by FC after grinding showed abnormal mature B cells, five cases were missed, all cases are not misdiagnosed. Among the specimens obtained by FNA, 18 cases showed FC-confirmed abnormal mature B cells, one case was missed, all cases are not misdiagnosed. The sensitivity, specificity, and accuracy of FC with CNB and FNA were 73.68 % (14/19) vs 94.73 % (18/19), 100 % (1/1) vs 100 % (1/1), and 75 % (15/20) vs 97.14 % (19/20), respectively. The sensitivity of the two puncture methods of FC of DLBCL was statistically different (p < 0.001). CONCLUSION: Sampling with ultrasound-guided FNA is of great value to improve the quality of FC specimens. FNA can significantly improve the sensitivity and accuracy of FC diagnosis in R/R DLBCL.

[Imatinib Combined with VP Low Dose Regiment for Treating Newly Diagnosed Adult Patients with Ph-positive ALL].

OBJECTIVE: To investigate the inductive therapeutic effects of imatinib combined with VP low dose regiment on adult patients with Ph-positive acute lymphoblastic leukemia (Ph(+) ALL). METHODS: Fourteen newly diagnosed adult patients with Ph(+) ALL were treated with VP regimen, and imatinib (400 mg/d) was added at the 8(th) day. VP regimen would be stopped when neutropenia lasted for 1 week or complicated with infection, fever, etc. Therapeutic effects were assessed by bone marrow morphology and quantitative analysis of BCR/ABL:ABL at the 28(th) - 33(rd) day. Patients could be treated with imatinib combined with chemotherapy for consolidation and maintenance therapy or were treated with allogeneic hematopoietic stem cell transplantation after complete remission. RESULTS: Fourteen cases obtained CR1 after first course of treatment, the median decline of BCR/ABA:ABL was 55.89 (10.25 -180.97) %; during the induction chemotherapy, pulmonary infection occurred in 3 patients, diarrhea in 1 patients, facial edema in 3 patients, however, all these patients were cured after symptomatic treatment, only 1 patient died of relapse after transplantation. CONCLUSION: In the period of tyrosine kinase inhibitor (TKI), inductive chemotherapy combined with imatinib and low dose VP can obtaine satisfactory CR rate and decrease the toxicity of the traditional drugs. It is suggested that TKI combined with VP regimen chemotherapy can achieve CR1 and make possible for allo-HSCT, from which patients can achieve the long-term survival.

[Autologous hematopoietic stem cell transplantation for patients with diffuse large B-cell lymphoma: a retrospective study].

The purpose of this study was to investigate the efficacy of autologous hematopoietic stem cell transplantation (auto-HSCT) for patients with diffuse large B-cell lymphoma (DLBCL), and analyse the factors influencing prognosis. The clinical data of 67 patients with DLBCL received auto-HSCT from 1996 to 2011 and cumulative overall survival (OS), progression-free survival (PFS), transplant-related mortality (TRM), and relapse rate were retrospectively analyzed. The results showed that the median follow-up time was 40 months after transplantation. Three-year cumulative OS and PFS were 70.6% and 66.4% respectively, 5-year cumulative OS and PFS were 70.6% and 63.8% respectively, and TRM was 7.2%. One-year and three-year cumulative relapse rate were 16.5% and 23.7% respectively. Univariate analysis revealed that the age and pre-transplant disease status were significantly associated with poor prognosis (P < 0.05). It is concluded that auto-HSCT is a safe and effective therapeutic option for patients with DLBCL, especially for the young patients or patients with better remission.

[Analysis of clinical characteristics and prognosis of 13 cases of acute erythroleukemia].

The aim of this study was to investigate the clinical characteristics and prognosis of acute erythroleukemia (AEL, AML-M6). The clinical features and results of morphologic, immunophenotypic, cytogenetic and molecular biologic detections were retrospectively analyzed in 13 cases of AEL from 305 acute leukemia patients hospitalized between October 2007 and October 2012. The results showed that the expression of erythroid and non-erythroid cells increased at the same time. The myeloid antigens mainly expressed CD13/CD33/CD117/CD34, while the erythroid antigens expressed Gly and CD71. The karyotypic detection indicated that there were 1 case with normal karyotype, 3 cases with simple karyotypic abnormality and 2 cases with complex karyotypic abnormality, the other cases were not detected. The molecular biological detection found that the poor prognosis gene existed in 5 cases [38.5% (5/13)], including 3 cases with MLL-MLL fusion gene, 1 case with MLL mutation, and 1 cases with NRAS gene mutation, the abnormal genes were not detected in remainder 8 cases. After chemotherapy with decitabine, the complete remission (CR) rate achieved 53.5% (7/13), partial remission (DR) rate achieved 15.4% (2/13). Finally, 8 patients received allo-HSCT, the median overall survival (OS) was 20.7 months, 3 year survival rate was 79%, 3 year disease-free survival rate was 78%. It is concluded that the acute erythroleukemia is a rare subtype of AML, which is transformed from MDS and has harmful genes and poor prognosis. Allo-HSCT and treatment with decitabine may enhance the survival rate of AEL.

[Clinical significance of rapid detecting bone marrow BCL2/IGH and BCL6/IGH fusion genes in patients with diffuse large B cell lymphoma by multiplex PCR].

Diffuse large B cell lymphoma (DLBCL) is the most common aggressive non-Hodgkin's lymphoma (NHL), characterized by great heterogeneity in clinical manifestations and molecular genetics. This study was aimed to explore the clinical significance of applying multiplex PCR to detect BCL2/IGH and BCL6/IGH fusion genes in DLBCL. Multiplex PCR was used to detect bone marrow samples from 80 cases of DLBCL. The results showed that 12 patients (15%) carried BCL2/IGH or BCL6/IGH fusion genes, BCL2/IGH was found in 6 patients (7.5%), and BCL6/IGH in another 6 patients (7.5%). The patients with different molecular markers displayed different clinical features and outcomes. It is concluded that multiple PCR is rapid and accurate method to identify gene abnormalities in DLBCL, but further studying a quantitative or semi-quantitative assay for the expression of fusion genes is needed.

[Application of multiplex nested RT-PCR to detecting 10 fusion genes related with MLL gene in myelodysplastic syndrome].

This study was aimed to investigate the clinical value of multiplex nested reverse transcription PCR (RT-PCR) in detecting MLL-related fusion genes in myelodysplastic syndrome (MDS). Ten MLL-related genes (dupMLL, MLL-ELL, MLL-ENL, MLL-AF6, MLL-AF9, MLL-AF10, MLL-AF17, MLL-CBP, MLL-AF1P, MLL-AF1Q) in 221 MDS cases were detected by multiplex nested RT-PCR. The results indicated that 20 patients were detected with positive result among 221 patients and the positive rate was 9.05%. The number of the positive cases and positive rates of the above mentioned 10 fusion genes were in order: 7 (3.16%), 2 (0.9%), 1 (0.45%), 1 (0.45%), 2 (0.9%), 2 (0.9%), 1 (0.45%), 2 (0.9%), 1 (0.45%), 1 (10.45%). It is concluded that the multiplex nested RT-PCR has been confirmed to be able to detect 10 fusion genes in MDS patients, which can provide important evidences for assessing diagnosis and treatment, and give related necessary information about minimal residual disease and its prognosis.

Effect of RhoC on hepatocellular carcinoma cell growth and related molecular mechanisms / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To clarify the role of RhoC in the growth of hepatocellular carcinoma cells and its molecular mechanism, so as to explore the molecular target of tumor cell growth.</p><p><b>METHODS</b>siRNA-RhoC plasmid was constructed and RhoC gene silencing the cell-line of hepatocellular carcinoma was setup. Cell growth was assessed by MTT assay. AgNORs staining was applied to determine cell proliferation. Plate cell clone test was conducted to examine the capacity of cell clone formation. FACS was adopted to measure the course of cell cycle and semi-quantitative RT-PCR was used to determine the expression of cell cycle proteins. In order to further determine the effect of RhoC expression on cell growth, a RhoC over-expression human hepatocellular cell line was setup by PcDNA3-RhoC plasmid transfection.</p><p><b>RESULTS</b>The inhibition rate of RhoC was 82.3%. From the fourth day of cell culture, the growth of cells in RNAi group was significantly slower than that in parental Bel7402 and negative control groups (0.41 ± 0.10 vs. 0.73 ± 0.11 and 0.71 ± 0.07 respectively, P < 0.05). AgNORs staining showed that average cell stained particles in RNAi group was significantly lower than that in parental Bel7402 and negative control(1.23 ± 0.35 vs. 3.47 ± 0.93 and 3.17 ± 0.78, P < 0.01). Plate clone formation test showed that clone formation efficiency in the RNAi group was notably lower than that in the control group [(20.33 ± 5.42)% vs. (70.58 ± 10.10)% and (69.83 ± 14.77)%, respectively, P < 0.01]. Cell cycle analysis by FACS showed that G(0)/G(1) cell percentage in the RNAi group was significantly higher than that in the control group [(73.14 ± 5.93)% vs. (57.05 ± 5.97)% and (52.99 ± 4.80)%, P < 0.05]. Compared with Bel7402 and negative control groups, the expression of following growth associated genes was significantly decreased: cyclin D1(0.45 ± 0.21 vs. 1.25 ± 0.24 and 1.12 ± 0.15, respectively, P < 0.05)and CDK4 (0.55 ± 0.08 vs. 1.18 ± 0.32 and 1.10 ± 0.29, respectively, P < 0.05); the following genes were notably increased: p16(1.07 ± 0.23 vs. 0.36 ± 0.12 and 0.35 ± 0.13, respectively, P < 0.01)and p21(0.42 ± 0.12 vs. 0.17 ± 0.06 and 0.19 ± 0.08, respectively, P < 0.05). RhoC was highly expressed in PcDNA3-RhoC transfected hepatocellular cell line. From the third day on of the cell culture, cell growth in PcDNA3-RhoC group was remarkably higher than that in the HL7702 and PcDNA3 groups (0.83 ± 0.10 vs. 0.54 ± 0.11 and 0.58 ± 0.55, respectively, P < 0.05).</p><p><b>CONCLUSIONS</b>RhoC is the key molecule in promoting hepatocellular cell growth, and is a promising target for tumor cell growth controlling.</p>

The value of spleen sub-pedicle two steps severance with LigaSure in laparoscopic splenectomy / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the safety and utility of the methods of spleen sub-pedicle two steps severance with LigaSure Vessel Sealing System combined with ultrasound scalpel to resect peri-splenic ligaments during laparoscopic splenectomy.</p><p><b>METHODS</b>The methods and the curative effect of 32 patients with laparoscopic splenectomy performed by the way as mentioned above were summarized and analyzed. 4 males, 28 females, median 36 years (range 16 - 64 years). Mean dimensions of spleens were 17 cm (range 11 - 23 cm). Nineteen patients had idiopathic thrombocytopenia purpura (ITP), 6 hereditary spherocytosis (HS) (5 of them were the same family constellation), 3 hemolytic anemia (HA), 2 spleen injury with haematoma infection, 1 Evan syndrome, 1 leukemia. Peri-splenic ligaments were resected by LigaSure combined with ultrasound scalpel, splenic pedicle was resected by spleen sub-pedicle two steps severance with LigaSure when splenic pedicle were sufficiently free. Splenic pedicle was non-excisional occluded by LigaSure at pancreatic cauda, in the first step and was occluded and resected alongside of the spleen in the second step.</p><p><b>RESULTS</b>All of the 32 cases were completely succeed, the average operating time was 70 min (55 - 130 min), the estimated intraoperative blood loss was 200 ml (50 - 600 ml), no complications such as abdominal haemorrhage, visceral injury, pancreatic leakage and infection were observed. All the patients were healed well, the average hospital stay after an operation was 6 days. The average expense of each patients could save 8050 yuan because Endo-GIA was not used.</p><p><b>CONCLUSIONS</b>Spleen sub-pedicle two steps severance with LigaSure Vessel Sealing System combined with ultrasound scalpel to resect peri-splenic ligaments during laparoscopic splenectomy, which shorten the operating persistence time with less hemorrhage is a safe, effective and low-cost minimally invasive surgery in selective cases.</p>