Plasma Quantitative Lipid Profiles: Identification of CarnitineC18:1-OH, CarnitineC18:2-OH and FFA (20:1) as Novel Biomarkers for Pre-warning and Prognosis in Acute Myocardial Infarction.

This study was aimed to determine the association between potential plasma lipid biomarkers and early screening and prognosis of Acute myocardial infarction (AMI). In the present study, a total of 795 differentially expressed lipid metabolites were detected based on ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Out of these metabolites, 25 lipid metabolites were identified which showed specifical expression in the AMI group compared with the healthy control (HC) group and unstable angina (UA) group. Then, we applied the least absolute shrinkage and selection operator (LASSO) and support vector machine-recursive feature elimination (SVM-RFE) methods to obtain three lipid molecules, including CarnitineC18:1-OH, CarnitineC18:2-OH and FFA (20:1). The three lipid metabolites and the diagnostic model exhibited well predictive ability in discriminating between AMI patients and UA patients in both the discovery and validation sets with an area under the curve (AUC) of 0.9. Univariate and multivariate logistic regression analyses indicated that the three lipid metabolites may serve as potential biomarkers for diagnosing AMI. A subsequent 1-year follow-up analysis indicated that the three lipid biomarkers also had prominent performance in predicting re-admission of patients with AMI due to cardiovascular events. In summary, we used quantitative lipid technology to delineate the characteristics of lipid metabolism in patients with AMI, and identified potential early diagnosis biomarkers of AMI via machine learning approach.

Construction of tissue-engineered lymphatic vessel using human adipose derived stem cells differentiated lymphatic endothelial like cells and decellularized arterial scaffold: A preliminary study.

We have previously demonstrated that human adipose-derived stem cells (hADSCs) can be differentiated into lymphatic endothelial like cells. The purpose of this study was to investigate the feasibility of utilizing the induced lymphatic endothelial like cells and decellularized arterial scaffold to construct the tissue-engineered lymphatic vessel. The hADSCs were isolated from adipose tissue in healthy adults and were characterized the multilineage differentiation potential. Decellularized arterial scaffold was prepared using the Triton x-100 method. ADSCs were differentiated into lymphatic-like endothelial cells, and the induced cells were then seeded onto the decellularized arterial scaffold to engineer the lymphatic vessel. The histological analyses were performed to examine the endothelialized construct. The decellularized arterial scaffold was successfully obtained and was able to maintain its vessel morphology. The isolated ADSCs can be differentiated into osteocytes and adipocytes. After seeding onto the scaffold, the seeded cells attached and grew well on the decellularized arterial scaffold. Our preliminary results demonstrated that the induced lymphatic endothelial like cells combined with decellularized arterial scaffold could be utilized to successfully engineer the lymphatic vessel. Our findings may be helpful for the development of tissue-engineering of the lymphatic graft.

In vitro induction of human adipose-derived stem cells into lymphatic endothelial-like cells.

Human adipose-derived stem cells (hADSCs) may provide a suitable number of progenitors for the treatment of lymphatic edema; however, to date the protocols for inducing hADSCs into this tissue type have not been standardized. We wished to investigate the induction of hADSCs into lymphatic endothelial-like cells using vascular endothelial growth factor-C156S (VEGF-C156S) and other growth factors in vitro. hADSCs from healthy adult adipose tissue were purified using enzyme digestion. Differentiation was induced using medium containing VEGF-C156S and bovine fibroblast growth factor (bFGF). Differentiation was confirmed using immunostaining for lymphatic vessel endothelial hyaluronan receptor (LYVE-1) and fms-related tyrosine kinase 4 (FLT-4), two lymphatic endothelial cell markers. The expression levels of LYVE-1, prospero homeobox 1 (PROX-1), and FLT-4 throughout induction were assessed using reverse transcriptase quantitative polymerase chain reaction. hADSCs were successfully obtained by trypsin digest and purification. Flow cytometry showed these cells were similar to mesenchymal stem cells, with a high positive rate of CD13, CD29, CD44, and CD105, and a low positive rate of CD31, CD34, CD45, and HLA-DR. Induction to lymphatic endothelial-like cells was successful, with cells expressing high levels of LYVE-1, PROX-1, and FLT-4. Adipose-derived stem cells can be induced to differentiate into lymphatic endothelial-like cells using a medium containing VEGF-C156S, bFGF, and other growth factors. This population of lymphatic endothelial-like cells may be useful for lymphatic reconstruction in the future.

Zoledronic acid exerts antitumor effects in NB4 acute promyelocytic leukemia cells by inducing apoptosis and S phase arrest.

The aim of this study was to investigate the antitumor effect of zoledronic acid (ZOL) in the NB4 human acute promyelocytic leukemia (APL) cell line and explore the potential mechanism of action of this compound. NB4 cells were exposed to various concentrations (0-200µM) of ZOL. Cell viability was measured by MTS assay. The extent of cell apoptosis and distribution of cells in the different phases of the cell cycle were analyzed with flow cytometry. The expression of apoptosis- and cell cycle-related proteins was assayed by Western blot. The combined effect of ZOL and arsenic trioxide (ATO) on the proliferation of NB4 cells was also determined. The results of this study indicate that ZOL inhibits cell proliferation in a time- and dose-dependent fashion and also induces apoptosis and S phase arrest in a dose-dependent manner. The Western blot analysis confirmed the induction of apoptosis and S phase arrest, revealing that the pro-apoptosis proteins Bax, Puma and activated caspase-9 were upregulated and the anti-apoptosis proteins Bcl-2 and Bcl-xL were downregulated. ZOL at a concentration of 50µM synergized with 0.5µM ATO on the growth inhibition of NB4 cells. Taken together, our data indicate that ZOL exerts a potent antitumor effect on NB4 cells by inducing apoptosis and cell cycle arrest, and that ZOL can synergize with the traditional chemotherapy drug ATO.

Prognostic value of miR-96 in patients with acute myeloid leukemia.

OBJECTIVE: Aberrant expression of miRNA (miR)-96 is associated with tumorigenesis and tumor progression in several solid cancers. However, little is known about the expression and prognostic value of miR-96 in acute myeloid leukemia (AML). Therefore, the aim of this study was to investigate the correlation of miR-96 expression with clinicopathological features and prognosis of AML. METHODS: Real-time quantitative RT-PCR assay was performed to evaluate the expression levels of miR-96 in mononuclear cells from bone marrow or peripheral blood specimens in 86 patients with newly diagnosed AML. RESULTS: Compared with normal controls, miR-96 expression was significantly downregulated in patients with newly diagnosed AML (P < 0.001). In analysis of 14 diagnosis/CR-paired samples, the expression level of miR-96 was found markedly elevated in patients after treatment than before (P < 0.001). Moreover, lower levels of miR-96 were associated with a higher white blood cell count, bone marrow blast count (P < 0.001 and 0.022, respectively), and lower hemoglobin and platelet count (P = 0.036 and 0.033, respectively). Although the low-expression group seemed to have a lower CR rate (53.85% vs 70.0%), there was no significant difference between the two groups (P = 0.213). The low-expression group had a lower relapse-free survival (RFS) (P = 0.038) and overall survival (OS) (P = 0.022) compared with the high-expression group during a median follow-up of 20 months. CONCLUSION: Our data demonstrated that the expression of miR-96 was downregulated in newly diagnosed AML patients and associated with leukemic burden, as well as RFS and OS. This suggests that miR-96 detection might become a potential biomarker of prognosis and monitoring in AML. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1434808553949498.

[Effects of human umbilical cord mesenchymal stem cells in the treatment of paraquat-induced lung injury].

OBJECTIVE: To evaluate the clinical effect and safety of human umbilical cord mesenchymal stem cells (HUCMSCs) in the treatment of lung injury caused by acute paraquat poisoning. METHODS: Thirteen patients with lung injury caused by acute paraquat poisoning, who were admitted to Guangzhou No. 12 People's Hospital from December 2008 to December 2012, were divided into HUCMSC group (n = 5) and control group (n = 8). All patients received conventional treatment, while the HUCMSC group was treated with HUCMSCs as an addition. Sequential Organ Failure Assessment (SOFA) system, which was created by the Infection Section of European Society of Intensive Care Medicine, and Acute Physiology and Chronic Health Evaluation II were used to acquire the SOFA scores of patients. The lung injury was evaluated with lung injury score (LIS). The two groups were compared with respect to maximum SOFA scores at 1, 3, 5, 7, 14, and 15 days after paraquat poisoning. RESULTS: The HUCMSC group showed significantly lower maximum SOFA scores than the control group at 15d after poisoning (1.80 ± 2.05 vs 13.50 ± 7.59, P < 0.05). The LISs of the HUCMSC group after treatment (0.45 ± 0.27) were significantly lower than those of the HUCMSC group before treatment (1.15 ± 0.34) and those of the control group after treatment (2.94 ± 1.20) (P < 0.01). In the HUCMSC group, all patients survived, and they complained no discomfort and showed normal liver, kidney, and lung functions in reexamination; one patient showed incompletely absorbed shadow in the posterior segment of the left lower lobe of the lung during lung CT scan, and no abnormal findings were seen in other patients. In the control group, one patient survived, and others died. No adverse reactions, such as chill and fever, were presented in the HUCMSC group. CONCLUSION: HUCMSCs show promise for clinical application in the treatment of lung injury caused by acute paraquat poisoning.

Analyzing gene expression profile in K562 cells exposed to sodium valproate using microarray combined with the connectivity map database.

To explore the mechanism underlying antileukaemia effect of sodium valproate, the growth and survival of the K562 cell line were investigated. Global profiles of gene expression in K562 cells exposed to sodium valproate were assessed and validated. The differentially expressed genes identified were further used to query the connectivity map database to retrieve a ranked list of compounds that act on the same intracellular targets as sodium valproate. A significant increase in cell apoptosis and a change in gene expression profile were observed in valproate-exposed K562 cells. The significant enrichment analysis of gene ontology terms for the differentially expressed genes showed that these genes were involved in many important biological processes. Eight differentially expressed genes involved in apoptosis were verified by quantitative real-time PCR. The connectivity map analysis showed gene expression profile in K562 cells exposed to sodium valproate was most similar to that of HDACi and PI3K inhibitors, suggesting that sodium valproate might exert antileukaemic action by inhibiting HDAC as well as inhibiting PI3K pathway. In conclusion, our data might provide clues to elucidate the molecular and therapeutic potential of VPA in leukaemia treatment, and the connectivity map is a useful tool for exploring the molecular mechanism of drug action.

Using an exon microarray to identify a global profile of gene expression and alternative splicing in K562 cells exposed to sodium valproate.

To investigate the effect of valproate treatment on the K562 cell line, a model for chronic myelogenous leukaemia, the growth and survival of the K562 cell line were investigated using the Annexin-V/PI dual staining method, and global profiles of gene expression and alternative splicing in K562 cells were assessed using exon microarrays. A significant increase in cell apoptosis was observed in valproate-exposed K562 cells using flow cytometry. A total of 628 transcripts were identified as being significantly differentially expressed. The number of genes demonstrating increased expression levels was greater than the number of genes demonstrating decreased expression levels (445 genes vs. 183 genes, respectively). The significant enrichment analysis of GO terms for the differentially expressed genes revealed that these genes are involved in many important biological processes such as apoptosis. Six of the genes observed to be differentially expressed that might be involved in apoptosis were selected to undergo qRT-PCR validation. In total, 198 candidates of alternative splicing variants were identified. Among them, three alternative splicing events were selected for validation, and CBLC and TBX1 were confirmed to be alternatively spliced by semi-nested PCR. In conclusion, valproate exposure facilitated cell apoptosis, altered mRNA expression and alternative splicing events in the K562 cell line.

Transplantation of adipose-derived stem cells overexpressing hHGF into cardiac tissue.

The delivery of adipose-derived stem cells (ADSCs) for promoting tissue repair has become a potential new therapy, while hepatocyte growth factor (HGF) is an important growth factor with angiogenic, antifibrotic, and anti-inflammatory benefits. Therefore, transplantation of ADSCs into acute myocardial infarction (AMI) may improve cardiac function through angiogenesis and anti-fibrosis, and that hHGF may enhance these effects. ADSCs were isolated from human subcutaneous adipose tissue. Lentivirus vector encoding human HGF (lenti-hHGF) was constructed and infected into ADSCs. Results indicated that transplantation of ADSCs led to improvement of left ventricular function, explained partly through their ability to differentiate into endothelial cells, resulting in increased blood flow and decreased fibrosis. Furthermore, hHGF enhanced these effects. This suggests that ADSCs combined with HGF gene transfer may be a useful strategy for the treatment of patients with ischemic heart disease.

15d-PGJ2 inhibits cell growth and induces apoptosis of MCG-803 human gastric cancer cell line.

AIM: To investigate the influence of peroxisome proliferator-activated receptor gamma (PPARgamma) ligand, 15-deoxy-12, 14-prostaglandin J2 (15dPGJ2) on the proliferation and apoptosis of MCG-803 human gastric cancer cell lines. METHODS: Cell proliferation was measured by 3H-TdR assay. Apoptosis was determined by ELISA and TUNEL staining. Protein and mRNA level of bcl-2 family and COXs were measured by Western blotting and Northern blotting respectively. PGE2 production was examined by RIA. RESULTS: 15dPGJ2 inhibited cell growth and induced apoptosis of MCG-803 cells. The COX-2 and bcl-2/bax ratios were decreased following 15dPGJ2 treatment. The PGE2 production in supernatants was also decreased. These changes were in a dose-dependent manner. CONCLUSION: 15dPGJ2 may be a useful therapeutic agent for the treatment of gastric cancer.

[Relationship of PTEN, Mdm2 and p53 expression in astrocytoma].

OBJECTIVE: To study the incidence of PTEN, Mdm2 and p53 expression in different histopathological grades of astrocytoma and the signal transduction mechanism through which PTEN affects Mdm2 and p53 expression. METHODS: Surgical specimens from 68 brain astrocytoma patients were analyzed to detect PTEN, Mdm2 and p53 expression by immunohistochemical method. RESULTS: The incidence of PTEN, Mdm2 and p53 expression was 54.4% (37/68), 41.2% (28/68) and 45.6% (31/68). The positive incidence of Mdm2 was 24.3% (9/37) in 37 cases with PTEN expression and 61.3% (19/31) in 31 cases without, which had a negative correlation between Mdm2 expression and PTEN expression (P < 0.01). Eighteen cases showed Mdm2(+)/p53(+) and 27 cases showed Mdm2(-)/p53(-). A significant correlation was found between Mdm2 and p53 expression (P < 0.05). CONCLUSION: The histopathological grade of astrocytoma is correlated with the loss of PTEN expression, Mdm2 amplification and p53 mutation. Mdm2 amplification, being in accordance with p53 mutation, can be inhibited by PTEN expression.

[Relationship between cyclins and prognosis of acute leukemia].

BACKGROUND & OBJECTIVE: Cell proliferation and differentiation are directed by cell cycle mechanism. When tumor cells proliferate abnormally, cyclins, which are positive agents of cell cycle, may be expressed abnormally at the same time. Now many references proved that cyclins are highly expressed in solid tumors. This study was designed to investigate the relationship between expression of cyclins and prognosis of acute leukemia. METHODS: Sixty-eight cases of acute leukemia were enrolled. Reverse transcription polymerase chain reaction (RT-PCR) was performed on tumor samples to examine the expression of cyclin A, cyclin D, cyclin E. All samples were divided into three groups: acute leukemia in complete remission (12 cases), newly diagnosed acute leukemia (16 cases) and refractory leukemia (40 case). Samples of benign hemopoietic patients were used as normal control (15 cases). RESULTS: All the 15 cases in the control group were negative of cyclin mRNA. No difference of cyclin A mRNA expression was shown between control group and the three experiment groups (P >0.05). But expression of cyclin D and cyclin E mRNA was significantly different between them (P< 0.01). There was no difference of expression of cyclin D and cyclin E mRNA among CR patients with acute leukemia(P >0.05), while the expression of cyclin D mRNA is significantly higher than that of cyclin E in refractory leukemia group. Furthermore the expression of cyclin D mRNA in recurrent refractory leukemia patients is significantly higher than that newly diagnosed cases (P< 0.05). All cyclins mRNA positive cases were divided into single cyclin gene expressed group and multiple cyclins gene expressed group. The positive rates of them were counted. No difference was found between complete remission group and refractory leukemia group (P >0.05). CONCLUSION: Cyclin D may act as a prognostic marker for acute leukemia. The amount of cyclins expressed cannot be used as a prognostic factor for acute leukemia.

[Relationship between tumor necrosis factor genetic polymorphisms and acute lymphocytic leukemia].

BACKGROUND & OBJECTIVE: Acute lymphocytic leukemia(ALL), which is sensitive to tumor necrosis factor (TNF) is one of hematological malignances derived from lymphoid tissue. Genetic polymorphisms in the tumor necrosis factor (TNF) locus can affect transcription and expression of TNF genes. This study was designed to investigate the relationship between -308 bp polymorphism in tumor necrosis factor-alpha(TNFalpha) gene and +252 bp in lymphotoxin-alpha(LTalpha) gene and the pathogenesis, clinical course, and outcome of ALL. METHODS: The single base mutation polymorphism in TNFalpha gene and LTalpha gene were analyzed among 29 Chinese patients with ALL and 72 normal controls using polymerase chain reaction (PCR)-restrictive fragment length polymorphism (RFLP). The clinical data were collected and survival analysis was performed. RESULTS: The difference of distribution of genotypes, alleles of TNFalpha(-308), LTalpha(+252), and TNF/LT polymorphic extended haplotypes between the ALL patients and control group were not statistically significant (P >0.05). In patients, no statistically significant association was found between the presence of a given TNF/LT haplotype status and clinical characters such as sex, white blood cell (WBC) counts, central nervous system involvement, and the response to therapy(P >0.05). The estimated 1-year overall survival rates in the groups of patients carrying high-risk and low-risk haplotypes were not statistically significant (P >0.05) using Kaplan-Meier method. In multivariate Cox regression models the TNF/LT haplotype status was not found to be a risk factor for outcome (P >0.05). CONCLUSION: These data suggest that genetic polymorphisms in the TNFalpha(-308) and LTalpha(+252) are not crucial in the pathogenesis, clinical course, and outcome of ALL patients.

[Review of relationship between tumor necrosis factor genetic polymorphism and hematological malignancies].

Patients with malignant tumor including hematological malignancies have high plasma levels of tumor necrosis factor (TNF) and excessive production of TNF is associated with polymorphic variation of TNF locus. TNF polymorphism affects TNF expression by affecting transcriptional regulation. The aim of this paper was to review the correlation between TNF polymorphism and hematological malignancies such as lymphoma, chronic lymphocytic leukemia, hairy cell leukemia, acute lymphocytic leukemia and multiple myeloma and to make preparation for further research of their relationship.