Overexpression of FGF2 delays the progression of osteonecrosis of the femoral head activating the PI3K/Akt signaling pathway.

BACKGROUND: The purpose of the current study was to explore the role and underlying mechanism of FGF-2 in dexamethasone (DEX)-induced apoptosis in MC3T3-E1 cells. METHODS: GSE21727 was downloaded from the Gene Expression Omnibus (GEO) database to identify the differentially expressed genes (DEGs) by the limma/R package. MC3T3-E1 cells were exposed to DEX at different concentrations (0, 10-8, 10-7, 10-6, 10-5 and 10-4 mol/L), and cell viability, flow cytometry and TUNEL assay were used to detect cell proliferation and apoptosis. An FGF-2-pcDNA3 plasmid (oe-FGF-2) was used to overexpress FGF-2, and western blotting was conducted to detect protein expression. RESULTS: We found that FGF-2 was downregulated in the DEX-treated group. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated that DEGs were associated with PI3K/Akt signaling pathway. DEX downregulated FGF-2 gene and protein expression, inhibited viability and induced MC3T3-E1 cell apoptosis. Overexpression of FGF-2 reversed DEX-induced apoptosis in MC3T3-E1 cells. FGF-2-mediated anti-apoptosis was impaired by inactivating the PI3K/AKT pathway with LY294002. Moreover, overexpression of FGF2 delayed the progression of DEX-induced osteonecrosis of the femoral head (ONFH) animal model by regulation PI3K/Akt signaling pathway. CONCLUSION: In conclusion, FGF-2 is effective at inhibiting DEX-induced MC3T3-E1 cell apoptosis through regulating PI3K/Akt signaling pathway.

[Effect of miR-100 on Migration of Rat Bone Marrow Mesenchymal Stem Cells].

OBJECTIVE: To explore the effect of miR-100 on the migration of rat bone marrow mesenchymal stem cells(rBMMSC). METHODS: The rBMMSC were isolated by density gradient centrifugation, cell surface epitopes of CD105, CD45, CD34, CD29 and CD44 were analyzed by flow cytometry. The rBMMSC were transfected with miR-100 mimic or inhibitor, then the expression of miR-100 in transfected cells was detected by real-time PCR. Migration test was used to observe the effect of miR-100 on cell migration ability. The secretion level of chemokine SDF-1 in culture supernatant of cells was quantitatively detected by ELISA. RESULTS: The isolated cells were identified as BMMSC. After rBMMSC were transfected with miR-100 mimic or inhibitor, as compared with control group,the expression of miR-100 in rBMMSC significantly increased or decreased respectively. In the migration experiment, the rBMMSC migration was significantly inhibited in the miR-100 mimic group (P<0.01), while the rBMMSC migration was significantly enhanced in the miR-100 inhibitor group (P<0.01). The concentration of SDF-1 in the supernatant of the miR-100 mimic group and the miR-100 inhibitor group did not change significantly compared with the control group (P>0.05). CONCLUSION: miR-100 can significantly inhibit the migration of rBMMSC, but not significantly correlated with the SDF-1.

Bimetallic Bi/Pt peroxidase mimic and its bioanalytical applications.

In this work, bimetallic Bi/Pt nanoparticles in bovine serum albumin biomolecular scaffold (BSA-Bi/PtNPs) were synthesized through a facile and green method. As compared with BSA-PtNPs, the BSA-Bi/PtNPs possess enhanced peroxidase-like catalytic activity. Moreover, the BSA-Bi/PtNPs are stable in harsh conditions such as high temperature, extreme pH environments, and high ionic strength, as well as in common biological matrixes. These prominent advantages enable the BSA-Bi/PtNPs to be applied to a wide range of fields. Bioassays, such as serum glucose detection, extracellular hydrogen peroxide (H2O2) monitor, and cancer cells labeling, have been realized with satisfying results. The linear range of glucose determination was from 1 to 100 µM and the limit of detection (LOD) was 0.2 µM. The H2O2 released from each MCF-7 cell after stimulation was calculated to be 2.66 × 10-16 mol/s. By utilizing folic acid as a recognition element, tumor cell could be readily distinguished by BSA-Bi/PtNPs and the LOD for MCF-7 cell detection was 90 cells.

[Genetic polymorphism of the killer cell immunoglobulin-like receptor genes in the Inner Mongolian population].

OBJECTIVE: To investigate the killer cell immunoglobulin-like receptor (KIR) gene frequencies and genotypes distributions in the Inner Mongolian population. METHODS: Ninety genomic DNA samples were extracted from blood samples of randomly chosen Mongolian individuals. Gene-specific PCR amplification was used to identify genes present or absent for 16 KIR loci. KIR genotype distributions were obtained and compared to that of 24 populations published in literatures using principal component analysis by SAS8.0 software. Genetic tree was constructed by the calculate Nei's genetic distance. RESULTS: (1) The frequency of KIR 2DL2, 2DS2 in Mongolian individual is higher than that in north Mongoloid and less than that in Caucasian. (2) Haplotype AA was identified in 37.78% of individuals, which is higher than that in north Mongoloid and lower than that in Caucasian. (3) Mongolian was considered between north Mongoloid and Caucasian by principal component and genetic tree analysis. CONCLUSION: Mongolian might be affected by the north Mongoloid and Caucasian, and showed intermediate between the two populations.

[Detection and clinical significance of JAK2 mutation in 412 patients with chronic myeloproliferative neoplasms].

OBJECTIVE: To investigate the frequency of JAK2V617F mutation in Chinese patients with chronic myeloproliferative neoplasms (MPN) and to study the relationship between JAK2V617F mutation and clinical characteristics. METHODS: JAK2V617F mutation was screened by allele-specific polymerase chain reaction (AS-PCR). RESULTS: JAK2V617F mutation was detected in 277 of the 412 patients with MPN. The frequency of JAK2V617F mutation was similar among essential thrombocythemia (ET), idiopathic myelofibrosis (IMF) and chronic myeloproliferative disorders-unclassified (MPD-U) (P > 0.05), but it was significantly lower than that in polycythemia vera (PV) (P < 0.05). The presence of JAK2V617F was found to be significantly correlative with advanced age at diagnosis (P < 0.01) and with higher hemoglobin levels and higher leukocyte counts (P < 0.05). Significant difference was found in complication of vascular events between JAK2V617 positive and negative patients (P < 0.05). JAK2V617F positive MPD-U patients were more prone to progress into typical MPN compared with JAK2V617F negative MPD-U patients. The association between abnormal karyotype and JAK2V617F was not found in cytogenetical analysis of 301 patients. CONCLUSION: The presence of JAK2V617F in MPD-U is associated with the disease development. There is a correlation between JAK2V617F mutation in MPN and advanced age, higher leukocyte counts, hemoglobin level and vascular events.

[Detection of JAK2V617F mutation and its clinical significance in 80 patients with M2 acute myelogenous leukemia].

OBJECTIVE: To explore the prevalence and prognostic significance of JAK2V617F gene mutation in acute myelogenous leukemia M2 (AML-M2) patients. METHODS: Allele specific polymerase chain reaction (PCR) was used to detect JAK2 gene mutation. RESULTS: Of 80 de novo AML-M2 patients, 6 at the time of first diagnosis and 1 at relapse were found to have JAK2V617F gene mutation (8.8%, 7/80). Morphologically, the whole blood and bone marrow of the 7 AML-M2 patients with JAK2V617F gene mutation presented a picture of acute leukemia instead of myeloproliferative disorders. Immunophenotypically, bone marrow samples showed myelogenous linage expression. Complete remission was obtained in 4 of 5 AML-M2 patients with JAK2V617F mutation who received treatment, while one patient had no response to the treatment. Follow-up was performed in all the 5 patients, with a median survival of 18.5 months in 4 patients. CONCLUSION: JAK2V617F gene mutation, as a type-1 mutation, might not be an initial event in the pathogenesis of acute myelogenous leukemia, and its presentation does not mean a poor prognosis in de novo AML patients.

[A quantitative assay for JAK2 mutation in 135 patients with chronic myeloproliferative neoplasms].

OBJECTIVE: To investigate the frequency and mutational status of JAK2 mutation in Chinese patients with chronic myeloproliferative neoplasms (MPN) and study the relative quantitation and clinical implications of mutated JAK2 transcript. METHODS: JAK2 mutation and the mutational status were screened with amplification-refractory mutation sequencing polymerase chain reaction (ARMS-PCR), the relative quantity of mutated JAK2 mRNA by using capillary electrophoresis. RESULTS: JAK2V617F mutation was detected in 95 of 135 MPN patients, including 37 (97.4%) of 38 polycythemia vera (PV), 56 (59.6%) of 94 essential thrombocythemia (ET) and 2 of 3 idiopathic myelofibrosis (IMF) patients; the difference between the mutations in PV and ET was significant (P<0.05). Of 95 JAK2V617F patients examined, 18/38 PV patients (47.3%) and 17/94 (18.1%) ET patients and 1 IMF patient were homozygotes, and ET patients showed lower prevalence of homozygote (P<0.05). In 95 MPN patients, the mutated mRNA ratio was higher in homozygote than in heterozygote patients. PV heterozygote patients showed higher levels of mutated JAK2 mRNA than ET heterozygote patients (P<0.05). The levels of JAK2V617F mRNA in patients over 60 years of age were significantly higher than that in those less than 60 years of age (P<0.001). Higher leukocyte counts were observed in PV and ET patients with higher levels of mutated JAK2 mRNA (P<0.05). The presence of JAK2V617F was found to be significantly associated with higher hemoglobin level in ET patients. Cytogenetic analysis was performed in 101 of the 135 patients, the association between abnormal karyotype and JAK2V617F was not found. CONCLUSION: The ARMS-PCR technique can be used to detect the frequency and mutational status of JAK2V617F mutation, and along with capillary electrophoresis, the estimation of minimal residual disease becomes possible.

[Osteotome sinus floor elevation without bone grafting: a 2-year prospective clinical study].

PURPOSE: The aim of this study was to evaluate the osteotome sinus floor elevation procedure with Straumann implants without bone grafting through observing the implant survival rate and the change of bone around the implant, and explored new bone formation and stability when the implants protruding in the sinus less than 3mm. METHODS: 23 patients with single upper maxillary molar lost were included in this study. The mean residual bone height (RBH) under the maxillary sinus was 5-8 mm and the distance of the implants protruding into the sinus was less than 3mm without bone grafting. A healing period of 3-4 months was allowed before abutment tightening at 35 N x cm. All the patients were followed up for 2 years to evaluate the implant survival rate. X-ray measurements (immediately after implant placement, after 3,6,12,24 months postoperatively) were performed to calculate the length of the implant protruding into the sinus and if the new bone formed near it. RESULTS: At the end of 2-year, the implant survival rate was 100% and the implants resisted the applied 35 N.cm torque and supported the definitive prosthesis well. The mesial and the distal sides of the implants protruding into the sinus had new bone formation. No hard tissue could be seen around the apical implant 3 months after inserting and the boundary between the new bone and the original sinus floor could not be seen 2 years after surgery. CONCLUSION: Elevation of the sinus membrane alone without addition of bone grafting could have the similar effect to the OSFE with bone grafting.

[Quantitative analysis for JAK2 mutation in 98 patients with essential thrombocythemia and its clinical significance].

The objective of this study was to identify the frequency and types of JAK2V617F mutation in chinese patients with essential thrombocythemia (ET), to quantitatively detect the level of mutation transcripts and to investigate its clinical significance. The frequency and types of JAK2V617F mutation were detected by amplification-refractory mutation sequencing polymerase chain reaction (ARMS-PCR), the transcript level of JAK2V617F mutation was determined by using capillary electrophoresis. The results indicated that the JAK2V617F mutation was detected in 59 out of 98 patient with ET, 18 of whom were homozygous mutation. The mean age of patients with homozygous and heterozygous mutation was higher than that of patients with wild type mutation (p < 0.05). The quantitative assay using capillary electrophoresis showed that the transcript level of JAK2V617F mutation in patients with homozygous mutation was (89.9 +/- 6.7)%, which was higher than that in patients with heterozygous mutation (57.1 +/- 6.7)% (p < 0.05); the transcript level of JAK2V617F mutation in patients with age < 60 years was (62.3 +/- 16.5)%, which was lower than that in patients with age > 60 years (72.4% +/- 15.8)% (p < 0.05). The rate of thrombotic complications in patients with JAK2V617F-positive was higher than that in patients with JAK2V617F-negative in which the rate of thrombotic complication in patients with homozygous mutation was higher than that in patients with heterozygous mutation (p < 0.05). Compared with patients without thrombotic events, there were higher level of transcripts of JAK2V617F mutation in patients with thrombotic events. It is concluded that the JAK2V617F positive and negative patients with ET display the different clinical features, therefore, the analysis of mutation types and detection of transcript levels not only helps to identify the disease status and progression, but also guides the treatment of ET patients.

[The quantitative assay and clinical significance of JAK2V617F mutation in 131 patients with chronic myeloproliferative disorders].

OBJECTIVE: To investigate the frequency and mutational status of JAK2V617F mutation in Chinese patients with chronic myeloproliferative disorders (CMPD) and to study the relative quantification of mutated JAK2 mRNA and the clinical significance. METHODS: JAK2V617F mutation and the mutational status were screened with amplification-refractory mutation system polymerase chain reaction (ARMS-PCR), the relative quantification of mutated JAK2 mRNA was studied by using capillary electrophoresis. RESULTS: A higher prevalence of JAK2V617F in either the heterozygote or homozygote status in essential thrombocythemia (ET) was observed in elderly patients with ET (P < 0.05). The presence of JAK2V617F was found to be significantly correlated with the age at diagnosis (P < 0.05); patients with age > or = 60 years showed significantly higher JAK2 mutated RNA levels than those with age < 60 years (P < 0.05); the presence of JAK2V617F in polycythemia vera (PV) and ET was found to be significantly associated with higher hemoglobin level and higher leukocyte count (P < 0.05). In addition, higher leukocyte count was observed in homozygous ET patients than in heterozygous ET patients (P < 0.05). The frequency of JAK2V617F mutation and the prevalence of homozygote in PV patients were higher than those in ET patients (P < 0.05). The differences of JAK2V617F mRNA levels among PV, ET and chronic idiopathic myelofibrosis (IMF) were not significant. CONCLUSIONS: ARMS-PCR technique can be used to detect the frequency and mutational status of JAK2V617F mutation owing to its sensitivity and along with capillary electrophoresis, quantitative assay for mutated JAK2 mRNA, diagnosis of CMPD and judgement of prognosis become possible.

[Expression of JAK2V617F and MPLW515L/K mutation in 30 suspected cases of early myeloproliferative disorders].

OBJECTIVE: To investigate the prevalence of JAK2V617F and MPLW515L/K mutation in patients with slightly elevated platelets (BPC) or hemoglobin (Hb) not meeting the criteria of polycythemia vera (PV) or essential thrombocythemia (ET). METHODS: Genomic DNA from bone marrow or blood mononuclear cells was screened with allele specific polymerase chain reaction (AS-PCR) for JAK2V617F and MPLW515L/K mutation. The history of thrombosis was assessed retrospectively by patients files. RESULTS: Of 30 patients, 14 (46.7%) were positive for the JAK2V617F mutation, none of them had the MPLW515L/ K. Five of these 14 patients had a history of thrombosis. Follow-up results were available in 22 patients. Among them, 12 patients with JAK2V617F mutation turned out to be MPD in 6-24 months; only 2 out of 10 patients without this mutation evolved to MPD. CONCLUSION: JAK2V617F mutation could be one of the diagnosis criteria of early MPD. No MPLW515L/K expression was found in early MPD.

[Interdigitating dendritic cell sarcoma-a case report with literature review].

OBJECTIVE: To report a case of interdigitating dendritic cell sarcoma (IDCS). PATIENT MATERIAL: The patient was a 41-year-old man with a lymph node bulging in the left neck. Laboratory examination of peripheral blood and bone marrow was abnormal. The diagnosis of IDCS was made by immunohistochemistry and electron microscopy. Treatment of this patient with ABVD regimen (adriamycin, bleomycin, vinblastine, dacarbazine) resulted in obvious improvement, but did not control the tumor infiltration. CONCLUSION: IDCS has no distinctive clinical or pathohistological characteristics. Immunohistochemistry and electron microscopy are crucial in distinguishing it from other histiocytic/dendritic cell neoplasms. IDCS displays an aggressive behaviour, and the responses to chemotherapy are variable.

[Cytogenetic feature of acute biphenotypic leukemia].

BACKGROUND & OBJECTIVE: Since the diagnostic criterion of acute biphenotypic leukemia(BAL) was recommended by European Group for the Immunological Characteristics of Leukemia (EGIL) in 1995, the reports on cytogenetic feature of BAL can be seen in homeland and outside, but the case numbers in the reports were low. This study was designed to explore the cytogenetic feature of BAL. METHODS: The FAB subsets of fifty-six BAL cases were established by morphology/cytochemistry; and surface immunophenotyping was performed by flow cytometry using a broad panel of lymphoid- and myeloid-associated monoclonal antibodies through directly immunofluorescence technique; karyotype analysis was carried out by reverse heating giemsa (RHG). RESULTS: The data from our study showed that the cytogenetic feature of BAL possess a certain degrees of heterogeneity. In all cases, 44.4% were normal karyotype, 55.6% were clonal chromosome aberration. In clonal chromosome aberration, Ph chromosome abnormality occurred 23.2% and in matched AML controls was 0%. Ph chromosome abnormality occurred 39.3% in group of coexpression of myeloid and B lymphoid lineage; Ph chromosome aberration occurred 0% in group of coexpression of myeloid and T-lymphoid lineage. The other associated clonal chromosome aberrations were t (8; 21), t (15; 17), t (9; 22), inv (16), and not associated clonal chromosome aberration were t (12; 17), t (14; 15), t (3; 6), +21, complex aberration, etc. CONCLUSION: The generalized abnormality of t (9; 22) in BAL indicate that the blast cells of BAL were derived from earlier hematopoietic stem/progenitor cell.