LncRNA CBR3-AS1 is associated with the BCR::ABL1 kinase independent mechanism of tyrosine kinase inhibitor (TKI) resistance in chronic myeloid leukemia patients.

Background: It is difficult for chronic myeloid leukemia (CML) patients with BCR::ABL1 independent drug resistance to achieve optimal efficacy. The aim of this study is to investigate the BCR::ABL1 kinase independent mechanism of tyrosine kinase inhibitor (TKI) resistance in CML patients to develop targeted therapeutic strategy. Methods: Herein, we analyzed the long non-coding RNA (lncRNA) and messenger RNA (mRNA) expression profiles of patients who achieved sustained deep molecular response (DMR) after TKI treatment and patients with non-DMR using RNA-seqencing. Furthermore, the differentially expressed lncRNAs and mRNAs were identified. The expression of chosen lncRNA was validated in an expanded cohort, and bioinformatics analysis was performed to analyze the function of selected mRNA. Results: LncRNA data analysis indicated the diversity lncRNA profiles among healthy individuals, CML patients with non-DMR, and CML patients with DMR. Differential expression analysis and Veen plot of up-regulated lncRNAs in patients with non-DMR (compared with healthy individuals) and down-regulated lncRNAs in patients with DMR (compared to patients with non-DMR) revealed that lncRNA CBR3-AS1 overexpression might be related to BCR::ABL1 independent TKI resistance of CML patients. The expression of CBR3-AS1 was then verified in an expanded cohort, suggesting that, compared with control group, there was no statistical difference of CBR3-AS1 expression in DMR group, whereas, CBR3-AS1 was up-regulated in non-DMR group. Moreover, the mRNA data analysis of RNA-sequencing was performed. We considered genes that up-regulated in non-DMR group (compared with control group), down-regulated in DMR group (compared with non-DMR group), showed no statistical difference between control and DMR group as the potential genes that associated with TKI resistance of CML patients. A total of 55 corresponding mRNAs were obtained including KCNA6, a target gene of CBR3-AS1. Further bioinformatics analysis showed that the major interacted genes of KCNA6 were enriched in several resistance-associated pathways including interleukin -17 signaling pathway and cyclic adenosine monophosphate signaling pathway. Conclusions: In conclusion, this work indicates that CBR3-AS1 might be involved in BCR::ABL1 independent TKI resistance of CML patients through targeting KCNA6, providing a novel target for intervention treatment of CML patients with BCR::ABL1 independent TKI resistance.

Cell division cycle 37 change after bortezomib-based induction therapy helps to predict clinical response and prognosis in multiple myeloma patients.

OBJECTIVE: Cell division cycle 37 (CDC37) modulates disease progression and bortezomib resistance in multiple myeloma by regulating X-box binding protein 1, nuclear factor-kappa-B, etc. This study aimed to explore the prognostic implication of CDC37 before and after bortezomib-based induction treatment in multiple myeloma patients. METHODS: CDC37 was detected from plasma cells of bone marrow by reverse transcription-quantitative polymerase chain reaction at baseline and after bortezomib-based induction treatment in 82 multiple myeloma patients, and in 20 disease controls and 20 healthy controls. RESULTS: CDC37 was increased in multiple myeloma patients versus disease controls and healthy controls (both P < 0.001). In multiple myeloma patients, CDC37 was related to increased serum creatinine (P = 0.017) and beta-2-microglobulin (P = 0.027), as well as unfavorable revised International Staging System stage (P = 0.041). Notably, CDC37 was reduced after bortezomib-based induction treatment versus that at baseline (P < 0.001). Furthermore, CDC37 at baseline was reduced in patients who achieved complete response versus those who did not achieve that (P = 0.023). Additionally, CDC37 after bortezomib-based induction treatment was also decreased in patients who achieved complete response (P < 0.001) and objective response (P = 0.001) versus those who did not reach them. Meanwhile, CDC37 at baseline only predicted worse progression-free survival (P = 0.033). Notably, CDC37 after bortezomib-based induction treatment estimated both shorter progression-free survival (P = 0.006) and overall survival (P = 0.005), which was confirmed by multivariate regression analysis. CONCLUSION: CDC37 decreases after bortezomib-based induction treatment, while its higher expression reflects unsatisfactory induction treatment response and survival in multiple myeloma.

Changes in Bacterial Communities and Their Effects on Soil Carbon Storage in Spartina alterniflora Invasion Areas, Coastal Wetland Bare Flats, and Sueada salsa Areas.

Spartina alterniflora is considered an invasive species that has affected the biogeochemical circle of carbon in coastal wetlands around the world. Nevertheless, it is still unclear how S. alternation invasion affects the carbon storage capacity of coastal wetlands as carbon pools through bacterial changes. Herein, bacterial communities and soil carbon content in coastal wetland native areas and S. alterniflora invasion areas were detected. It was found that an S. alterniflora invasion brought more organic carbon and resulted in the increase in Proteobacteria in bare flats and Sueada salsa areas. When decomposition capacity was not sufficient, large amounts of organic carbon may be stored in specific chemical forms, such as monosaccharides, carboxylic acids, alcohols, etc. The results have also shown that soil bacterial communities were highly similar between the bare flat and S. alterniflora invasion area, which is extremely conducive to the rapid growth of S. alterniflora. However, an S. alterniflora invasion would decrease total carbon contents and inorganic carbon contents in the Sueada salsa area. This is not conducive to the stability of the soil carbon pool and soil health. These findings may complement, to some extent, the shortcomings of the interaction between S. alterniflora and bacterial communities, and their joint effect on soil carbon storage.

Insights from a patient with chronic lymphocytic leukemia complicating ALK+ anaplastic large cell lymphoma.

Chronic lymphocytic leukemia (CLL) that transforms into a more aggressive lymphoma has been termed Richter syndrome (RS). CLL with T-cell neoplasia is rarely reported; those with ALK+ anaplastic large cell lymphoma (ALCL) are also exceedingly rarely reported. A 63-year-old woman from the south of China presented with generalized lymphadenectasis and fever; she already had a prior diagnosis of CLL 9 years ago. As per her current diagnosis, it was CLL with ALK+ ALCL. The two-lymph node and bone marrow biopsies presented two types of cellular groups: i) left cervical lymph node biopsy suggested CLL (Ki67: 10%), along with bone marrow biopsy exhibited enhancement of the small lymphocytes (30%) with scant cytoplasm, round or irregular cell nuclei, and massive amounts of chromatin. Large cells (< 1%) that expressed CD30 and ALK were visible; The results of immunohistochemistry were as follows: CD20 (weak positive); PAX5 (positive); CD23 and CD5 (weak positive); and CD3, CD10, and CyclinD1 (negative); ii) left supraclavicular lymph node biopsy suggested ALK+ ALCL (Ki67: 70%). The final diagnosis was CLL with ALCL. The mechanisms of this condition are not fully understood, which might be associated with chronic stimulation of T cells by CLL cells along with immune dysfunction.

A promising strategy for the treatment of ischemic heart disease: Mesenchymal stem cell-mediated vascular endothelial growth factor gene transfer in rats.

BACKGROUND: Treatment of ischemic heart disease (IHD) remains a worldwide problem. Gene therapy, and recently, cell transplantation, have made desirable progress. A combination of appropriate stem cells and angiogenic genes appears promising in treating IHD. OBJECTIVE: To study the results of angiogenesis and myogenesis induced by transplantation of the adenovirus carrying human vascular endothelial growth factor 165 (Ad-hVEGF(165))-transfected mesenchymal stem cells (MSCs) in IHD compared with direct MSC transplantation or Ad-hVEGF165 delivery. METHODS: Cultured MSCs were transfected by Ad-hVEGF(165), and secreted VEGF was measured by ELISA in vitro. Ad-hVEGF(165)-transfected MSCs (MSC/VEGF group), MSCs (MSC group), Ad-hVEGF(165) (VEGF group) or a serum-free medium (control group) was injected into syngeneic Wistar rats immediately after left coronary artery occlusion. All cells were marked with CM-DiI (Molecular Probes, USA) before transplantation. One week after treatment, messenger RNA expression of hVEGF(165) in the MSC/VEGF group was found to be significantly higher than in other groups by reverse transcriptase-polymerase chain reaction analysis. One month after cell transplantation, left ventricular (LV) ejection fraction, capillary density of the infarcted region, infarct size and hemodynamic parameters (including LV end-diastolic pressure, LV+dP/dt and LV-dP/dt) were measured and immunohistochemical analysis was performed. RESULTS: A high level of VEGF was expressed by Ad-hVEGF(165)-transfected MSCs. LV ejection fraction, mean capillary density of the infarcted region and hemodynamic parameters were significantly improved in the MSC/VEGF group compared with the MSC group, the VEGF group and the control group (P<0.001 for all). Partly transplanted MSCs showed the cardiomyocyte phenotype, expressed desmin and cardiac troponin T, and resulted in angiogenesis in the ischemic myocardium. However, a few transplanted MSCs incorporated into the vascular structure and most of the new vascular components were host-derived. CONCLUSIONS: The combined strategy of MSC transplantation and VEGF gene therapy can produce effective myogenesis and host-derived angiogenesis, resulting in the prevention of progressive heart dysfunction after myocardial infarction.

[Reoperation about recurrent heart valve disease in 221 cases].

OBJECTIVE: To retrospectively review the experience of reoperation after closed mitral commissurotomy, valvuloplasty, perivalvular leakage and dysfunction of bioprosthetic valve in 221 cases. METHODS: Two hundred and twenty-one patients underwent heart valve reoperation from January 1998 to August 2005. Among them, 8 cases was emergency operation. The reasons of reoperation included 105 cases suffered from mitral valve restenosis after closed mitral commisurotomy, 37 cases suffered from valve lesion after mitral or aortic valvuloplasty, 29 cases suffered from perivalvular leakage after valve replacement. Eighteen cases suffered from bioprosthetic valve decline, 9 cases suffered from dysfunction of machine valve, 7 cases suffered from tricuspid insufficiency of Ebstein, 5 cases suffered from prosthetic valve endocarditis and 11 cases suffered from other valve disease. The re-operations were mitral valve replacement, mitral and aortic valve replacement, aortic valve replacement and tricuspid valve replacement. The interval from first operation to next operation was 1 - 21 years. RESULTS: The early-stage postoperative mortality was 8.6% (19/221). And the reasons were low cardiac output syndrome, arrhythmia, multiple organ dysfunction failure (MODF) and renal failure. Among these the emergency operative mortality was 3/8. And the mortality was 14.5% (9/62) in class IV of cardiac function (NYHA). CONCLUSIONS: The risk factors of reoperation about heart valve disease include emergency operation, low preoperative cardiac function, MODF, long time of cardiopulmonary bypass and aortic blocking. Therefore it is emphasized that mastering and treating the risk factors promptly, which could decrease the mortality and incidence of complication.

[Experimental study of differentiation of mouse bone marrow stromal stem cells into progenitor cardiomyocyte in vitro].

OBJECTIVE: To investigate the possibility of inducing mouse bone marrow stromal stem cells (MSCs) into progenitor cardiomyocytes in vitro. METHODS: The MSCs were isolated by adhesion culture in vitro and flow cytometery was employed to identify the phenotypes of the cell passages. 5-azacytidine was used to induce the stem cells to differentiate into cardiomyotes in the experimental group, which were then detected by RT-PCR, semi-quantitative RT-PCR, Western-blot analysis, electron microscopy and immunofluorescence technique. RESULTS: The isolated subcultured MSCs displayed a fusiform cell-like morphology. The MSCs were uniformly positive for CD29 and CD44 but negative for CD34 and CD45, and after induction, they expressed cardiomyocyte-specific transcription factors (NKx2-5/Csx and GATA4) and fetal ventricular cardiomyocyte-specific gene beta-myosin heavy chain, but not adult ventricular cardiomyocyte-specific gene alpha-myosin heavy chain as detected by RT-PCR. Western blotting identified the expressions of alpha-sarcomeric actin and desmin in the induced MSCs, with myofilament formation observed under electron microscope. Compared with the control group, the expression level of DNA methyltransferase mRNA was significantly lowered in the experimental group as observed by semi-quantitative RT-PCR (P<0.05). CONCLUSION: MSCs are capable of differentiating into progenitor cardiomyocytes.

[Fontan operation with extracardiac conduit on beating hearts].

OBJECTIVE: To evaluate the results of Fontan operation with extracardiac conduit on beating hearts. METHODS: Forty-two patients (31 males and 11 females) age ranged from 3 to 19 years old included in this study. There were 19 double inlet-ventricle, 10 tricuspid atresia, and 3 patients with mitral atresia, 10 patients with other complex congenital cardiac malformations. Fontan operations with extracardiac conduit were performed in all patients with the help of cardiopulmonary bypass without hypothermia in this study. Atrial septal fenestration was performed in 8 patients. In one patient, bi-directional cardiopulmonary procedure was performed 2 years before Fontan operation. RESULTS: There was one early death caused by acute hepatic function failure and one late death caused by repeated lung infections. The follow-up of 1 to 4.5 years showed that all patients' cardiac functions were grade I to II, and arterial oxygen saturation was 92% - 96%. CONCLUSIONS: The early and mid-term outcome of Fontan operation with extracardiac conduit on beating hearts is good and the method can be used in the single ventricle repair.