AIM:To study the morphological changes of cardiac H9c2 cells during the developmental process of fetal rat.METHODS:Embryonic rat heart-derived H9c2 cells were maintained in DMEM supplemented with 10%fetal bovine serum.The H9c2 cells were plated at a density of 6 000 cells/cm and divided into 5 groups:H9c2 cells were trea-ted with 5 mmol/L glucose, 25 mmol/L glucose, 50 mmol/L glucose, Norvasc (25 nmol/L) +25 mmol/L glucose, or Norvasc (25 nmol/L)+50 mmol/L glucose for 48 h.The morphology of H9c2 cells was observed.The cell surface area was measured by Image-Pro Plus 6.1 software.Fluorescence spectrophotometry was used to detect the concentration of in-tracellular calcium ion ( [ Ca2+] i ) in the cardiomyocytes.The concentration of CaN in the cell was measured by ELISA. The mRNA expression of CaNAβ, NFAT3 and β-MHC in the cells was detected by real-time PCR.The protein levels of CaNAβ, NFAT3 and β-MHC in cultural H9c2 cells were detected by Western blot.RESULTS: The mean area of the cells, the mean fluorescence value of [ Ca2+] i and the concentration of CaN in 25 mmol/L glucose group were higher than those in 5 mmol/L glucose group, and those were lower than those in 50 mmol/L glucose group.After treated with Nor-vasc, those results decreased significantly.The expression of CaNAβ, NFAT3 andβ-MHC at mRNA and protein levels in 25 mmol/L glucose group was higher than those in 5 mmol/L glucose group, but was lower than those in 50 mmol/L glu-cose group .The expression of CaNAβ, NFAT3 andβ-MHC at mRNA and protein levels decreased significantly in Norvasc treatment group.CONCLUSION:Ca2+-CaN-NFAT3 signaling pathway is perhaps involved in high glucose-induced H9c2 cardiomyocyte hypertrophy.

AIM: To investigate the effects of octreotide on metabolism in the A549 cells treated with lipopo-lysaccharides ( LPS).METHODS: The technologies of gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS) were used to test the metabolism of lung A549 cells subject to different treat-ment with LPS and/or octreotide.The results were visualized and checked by chromatogram, and the corresponding intensi-ty data were analyzed by the principal component analysis(PCA)method.The metabolites with different expression and the underlying interaction network were resolved.RESULTS: The metabolism analysis by LC/MS method indicated that there were different expression levels between different treated groups.Further analysis was carried out by orthogonal projections to latent structures-discriminant analysis (OPLS-DA) and the different expressed metabolites were obtained, which were mainly amino acids and phospholipids.By analyzing with GC/MS method and t-test, the different expressed metabolites were mainly organic acid, saccharides and amino acid metabolite.The interaction network diagram was constructed about the response of A549 cells induced by LPS and/or octreotide, including glycolysis/gluconenogenesis, tryptophan metabo-lism, galactose metabolism, urine cycle and citrate cycle.Fourteen key components were found such as serotonin, indole, threonine, serine, glucose, phenylalanine, lactose, fumarate, 4-hydroxyphenyllactate, aspartate, asparagine, putrescine, proline and succinate.CONCLUSION: In octreotide treated LPS-induced A549 cells, the main metabolites are organic acid, saccharides, amino acids and phospholipids.The interaction network is constructed, including 5 metabolic pathways
and 14 key components.

OBJECTIVETo compare the clinical effects of two methods of internal fixation in treating unstable femoral intertrochanteric fractures in elderly patients.

METHODSFrom August 2009 to August 2014, 68 elderly patients with unstable femoral intertrochanteric fracture treated with locking proximal femur plate and auxiliary short reconstructed plate (reconstructing calcar group) and proximal femoral nail antirotation (PFNA group) with clinical course from 1 to 3 days were retrospectively analyzed. In reconstructing calcar group, there were 30 patients including 8 males and 22 females, aged from 63 to 85 years old with an average of (73.41±5.12) years old, the fractures were classified to type AO 31-A2.2 in 12 cases, A2.3 in 11 cases, A3.3 in 7 cases according to AO/ASIF classification. In PFNA group, there were 38 patients including 10 males and 28 females, aged from 65 to 90 years old with an average of (74.26±4.53) years old, the fractures were classified to type AO 31-A2.2 in 15 cases, A2.3 in 13 cases, A3.3 in 10 cases. All fracture were caused by injury, leading pain and swelling. Femoral intertrochanteric fracture was confirmed by X ray films. The data of each group were collected for statistical analysis on the following aspects: the incision length, operation time, blood loss volume, postoperative partial weight bearing standing time, clinical healing time of fracture, postoperative complications, and hip functional score of Harris.

RESULTSAll incisions were healed at stage I. In the aspect of postoperative complications, there were 1 case of screw blade cutting and 1 case of deep venous thrombosis in PFNA group; there was 1 case of deep venous thrombosis in the reconstructing calcar group (²=0.000,=1.000). Patients were followed up from 20 to 24 months with an average of 22.5 months. There were no significant in postoperative partial weight bearing standing time, postoperative complications, hip functional score of Harris between two group. There were significant in the incision length, operation time, blood loss volume, clinical healing time of fracture. In the incision length, operation time, blood loss volume, clinical healing time of fracture, the PFNA group was significantly differently less than that of the reconstructing calcar group (<0.001). In the clinical healing time of fracture, the PFNA group was significantly differently less than that of the reconstructing calcar group (<0.05).

CONCLUSIONSFor the treatment of unstable femoral intertrochanteric fractures in elderly patients, reconstructing calcar and PFNA are both effective, and proximal femoral intramedullary nails may be the best choice, which can be simpler operation, smaller incision and less healing time.

The aim of this study was to investigate the expression of osteopontin (OPN) and vascular endothelial growth factor (VEGF) in acute leukemia (AL) and its significance in the angiogenesis and progress of AL. Serum levels of OPN and VEGF in 25 de novo patients, 19 complete remitted (CR) patients, 14 unremitted patients, and 11 relapsed patients with AL were measured by enzyme-linked immunosorbent assay (ELISA), and compared with those in normal controls. The results showed that the serum levels of OPN and VEGF in de novo, unremitted, relapsed patients were significantly higher than those in normal controls and CR patients (p < 0.01). Compared with de novo AL patients group, the serum levels of OPN and VEGF in CR patients decreased significantly, but showed no significant difference from those in normal controls (p > 0.05). The expression of OPN and VEGF in acute leukemia was positively correlated with occurrence and development of AL. It is concluded that the expressions of OPN and VEGF are closely related with AL occurrence and development, the OPN may regulate VEGF expression and promote angiogenesis in acute leukemia. Preventing expressions of OPN may seem as targets for leukemia therapy in the future.
Acute Disease ; Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Humans ; Leukemia ; blood ; pathology ; Male ; Middle Aged ; Neovascularization, Pathologic ; pathology ; Osteopontin ; blood ; Vascular Endothelial Growth Factor A ; blood ; Young Adult

This study was aimed to investigate the influence and significance of celecoxib (specific inhibitor of cyclooxygenase-2) on mRNA expressions of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (b-FGF), transforming growth factor β (TGF-β) in acute leukemia cells. The expressions of VEGF, b-FGF, TGF-β mRNA were measured by RT-PCR in acute leukemia cells treated with celecoxib (80 µmol/L, for 48 h) or with PBS. The results showed that the obvious expressions of VEGF, b-FGF, TGF-β mRNA were observed in acute leukemia cells. By using Pearson correlation analysis, there was positive correlation between VEGF mRNA and b-FGF mRNA expressions (r = 0.559, P = 0.001), and negative correlation between VEGF and TGF-β mRNA expressions (r = -0.4, P = 0.029). Expression levels of VEGF, b-FGF, TGF-β mRNA in experimental group were lower than that in control group (P < 0.01). It is concluded that COX-2 inhibitor celecoxib can inhabit vascular endothelial growth through down-regulating the mRNA expression of VEGF, b-FGF and TGF-β in acute leukemia cells. COX-2 inhibitor may offer supplemental effect for treating acute leukemia.
Adult ; Celecoxib ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Humans ; Leukemia ; drug therapy ; metabolism ; Male ; Pyrazoles ; pharmacology ; RNA, Messenger ; genetics ; Signal Transduction ; Sulfonamides ; pharmacology ; Transforming Growth Factor beta ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

The aim of study was to investigate the expression of angiogenin-2 (Ang-2) and vascular endothelial growth factor (VEGF) expression in acute leukemia (AL) and its significance in the angiogenesis and progress of AL. Serum levels of Ang-2 and VEGF in 24 de novo, 18 complete remitted (CR), 7 unremitted, 13 relapsed patients with AL were measured by enzyme-linked immunosorbent assay (ELISA), and compared with those of normal controls. The results showed that the serum levels of Ang-2 and VEGF in de novo, unremitted and relapsed patients were significantly higher than those in normal controls (p < 0.01). Compared with de novo group, the serum levels of Ang-2 and VEGF in CR patients decreased significantly, but showing no significant difference from those in normal controls (p > 0.05). In relapsed patients, the serum levels of Ang-2 and VEGF were obviously higher than those in unremitted and CR patients (p < 0.01). It is concluded that the expressions of Ang-2 and VEGF are closely related with occurrence and development of acute leukemia, the VEGF may regulate Ang-2 expression and promote angiogenesis and acute leukemia development. Preventing expressions of Ang-2 and VEGF may seem as targets for leukemia therapy in the future.
Acute Disease ; Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Male ; Middle Aged ; Neovascularization, Pathologic ; pathology ; Ribonuclease, Pancreatic ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

OBJECTIVETo investigate the clinical efficacy of regimen consisting of lenalidomide combined with chemotherapy for acute leukemia and its impact on vascular endothilial growth factor (vEGF) and basic fibroblast growth factor (bFGF), and to analyze the relationship lenalidomide with therapeutic efficacy of leukemia.

METHODSThe patients with newly diagnosed acute myeloid leukemia (except M3) from October 2013 to October 2014 in our hospital were randomly divided into 2 groups: chemotherapy+placebo (CP) group and lenalidomide+chemotherapy (LC) group. In addition, healthy persons were used as healthy controls (HC). The expression of VEGF and bFGF was detected by ELISA, and the therapeutic efficacy for AML patients was analyzed.

RESULTSThe therapeutic efficacy in LC group and CP group was 87.9% and 77.2% respectively. Before treatment, the VEGF level in LC and CP groups was obviously higher than that in HC group; after treatment, the VEGF level significanthy decreased, and the decreased degree in LC group was larger than that in CP group. Before treatment, the bFGF level in LC and CP groups was higher than that in HC group; after treatment, the bFGF level decreased, and decreased degree in LC group was larger than that in CP group.

CONCLUSIONThe lenalidomide combined with chemotherapy can significantly decrease the expression level of VEGF and bFGF, and enhance the remission rate of patients with AML.

Acute Disease ; Antineoplastic Agents ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Fibroblast Growth Factor 2 ; metabolism ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Thalidomide ; administration & dosage ; analogs & derivatives ; therapeutic use ; Vascular Endothelial Growth Factor A ; metabolism

Objective: To isolate and identify Prevotella nigrescens (P.nigrescens) in gingival crevicular fluid of patients with chronic periodontitis and to analyze its tumor-promoting role in esophageal squamous cell carcinoma (ESCC). Methods: Samples of gingival crevicular fluid were collected from patients with chronic periodontitis and cultured on GAM agar medium under anaerobic conditions. Black colonies on GAM agar plates were picked for subculture, and then the bacteria were isolated and purified. Bacterial identification was conducted using Gram staining and 16S rDNA sequencing analysis. The isolated bacterial species was used to infect ESCC NE6-T cells and the changes in biological characteristics of the infected cells were evaluated. Results: Multiple bacterial colonies were observed after anaerobic culturing of gingival crevicular fluid samples for 120 h on GAM agar plates. A single bacterial colony with pure black and smooth appearance was obtained from grey black bacterial colonies with streak method. It was a Gram-negative bacterium with bead-like shape. It showed 99.78% 16S rDNA sequence identity with P. nigrescens F0103 and was named P. nigrescens LY01. P. nigrescens LY01 infection promoted the in vitro proliferation, migration and invasion of NE6-T cells and the growth of subcutaneous xenograft in nude mice. In addition, it could also induce the upregulation of Ki67 and activation of p-STAT3. Conclusions P. nigrescens inhabiting in gingival sulcus might promote the progression of ESCC in human with chronic periodontitis.