Objective · To investigate the histone methyltransferase capability of DOT1L-long form and its role in breast cancer metastasis.Methods · The existence of DOT1L-long form was confirmed by PCR, and the mRNA level of DOT1L was tested by real-time PCR. In HEK293T cells in which DOT1L canonical and DOT1L-long were overexpressed respectively, Western blotting was used to test the expression level of DOT1L and the histone methyltransferase capability. In the MCF10A cell line with inducible expression of DOT1L-long, real-time PCR was used to detect the mRNA level of epithelial-mesenchymal transition (EMT) marker, and transwell assay was used to detect the migration of breast cancer cells in which the expression level of DOT1L is low or high. Results · PCR demonstrated the existence of DOT1L-long form, and real-time PCR showed it widely exists in HCT116, T98G, MCF10A cells, etc. Western blotting showed the expression of DOT1L-long form and its H3K79 methyltransferase activity. In MCF10A cells in which overexpressed canonical DOT1L and DOT1L-long, mRNA levels of N-cadherin and fibronectine increased. Transwell showed canonical DOT1L and DOT1L-long both substantially increased the migration of breast cancer cells. Conclusion · The existence of DOT1L-long was confirmed and investigated, which is 202 amino acids longer than the canonical DOT1L, and is coded by a new exon, located between exon 27 and 28. Further, the DOT1L-long has H3K79 methyltransferase activity, and is able to promote breast cancer metastasis.

OBJECTIVETo study the combination of Mandibular distraction and orthognathic techniques for the reconstruction of adult hemifacial microsomia.

METHODSThe three-dimensional CT reconstruction data was used with Mimics for preoperation design. The osteotomy location, distraction vector, distraction distance were decided before operation with a surgical guider. At the first stage, internal distractor was implanted after ostetomy through an extra-oral approach. The distraction begun 5-7 days after operation with a frequency of 1 mm/day. After distraction, the distractor was maintained for 3-6 months. At the second stage, the distractor was removed. Le Fort I osteotomy was performed in order to correct the cross-bite and improve the facial contour. Usually, bone graft was inserted into the gap after Le Fort I osteotomy. The genioplasty was also performed if necessary.

RESULTS9 cases of adult hemifacial microsomia with severe mandibular deviation were treated. The facial asymmetry were improved greatly. 1 patient suffered an wound infection in the maxillary region after Le Fort I osteotomy and healed uneventfully with wound irrigation.

CONCLUSIONSMandibular distraction combined with orthognathic surgery is an effective procedure for adult hemifacial microsomia with complicated mandibular hypoplasia.

Adult ; Aged ; Bone Transplantation ; Facial Asymmetry ; surgery ; Goldenhar Syndrome ; surgery ; Humans ; Mandible ; surgery ; Osteogenesis, Distraction ; methods ; Osteotomy, Le Fort ; methods

0.05), but the level of albumin dropped significantly(P

Antibiotics, Antineoplastic ; pharmacology ; Breast Neoplasms ; enzymology ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Female ; Glucosyltransferases ; biosynthesis ; genetics ; Humans ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

Objective To study the effect of platelet-derived growth factor(PDGF)and lysosomes on lung injury in macaque with early-phase endotoxie shock.Method Eleven macaques were randomly divided into two groups,namely,control group(Co group,n=5)iand endotoxic group(En group,n=6).The macaque of the Co group injected with 1 ml/kg normal saline and the macque of the En group received a dose of 2.8 mg/kg Lipopolysaccharides(LPS)i.v.The blood gas was detected at 120 minutes after LPS challenging. Uhrastructure,cytochemistry of acid phosphatase(ACPase)detection by electronic microscopy and immunohistochemical assay of PDGF were completed in hmgs of all the macaque .Results Administration of LPS did not change the parameters of gas exchange,namely,PaO_2,PaO_2/Fi and PaCO_2.In the early phase,of endotoxic shock,ACPase activity products increased and lysosome destroyed in the alveolar cells.The pathologic changes of alveolus,such as degeneration of vessel endothelium,injury of alveolar epithelium and damage of basement membrane,and transudation of blood component were observed by electron microscopy in the En group. However,no pathological changes were found in the control group.By immunohistochemical staining,PDGF on alveolar wall in the En animals was observed,whereas no PDGF protein in the Co macaques was noticed. Conclusions Administration of LPS induced the expression of PDGF in the alveolar wall and lysosome injury in the alveolar cells,as a result of alveolar damage in early-phase endotoxin shock.In the meantime,the parameters of gas exchanges did not change.The PDGF may play an important role in the pathogenesis of lung during the early-phase of endotoxin shock.

OBJECTIVETo investigate the effects of Gastric bypass surgery on the apoptosis of islet β-cells in type 2 nonobese diabetic (NOD) rats and its mechanisms.

METHODSSeventy-two 8-week-old GK rats were randomly divided into four groups:operation group (group O, n = 18), sham operation group (group S, n = 18), diet control group (group F, n = 18) and control group (group C, n = 18). The levels of fasting, postprandial blood glucose, insulin and glucagon-like peptide-1 (GLP-1) were measured and compared among the 4 groups before the operation and at 1, 2, 4 and 8 weeks following the operation. The blood samples were collected at 2, 4 and 8 weeks after the operation for the measurement of postprandial blood glucose, and then the rats in batches (6 rats in each group) were decapitated to retrieve the pancreas. The apoptosis of the islet β-cells was detected by using TUNEL assay, and the expression of apoptosis-related proteins Bcl-2, Bax was measured with immunohistochemistry.

RESULTSAs for group O, the fasting blood glucose level decreased from (16.2 ± 0.8) mmol/L before the operation to respectively (9.2 ± 0.6) mmol/L and (9.7 ± 0.7) mmol/L at 4 and 8 weeks after the operation; postprandial blood glucose decreased from (31.1 ± 1.1) mmol/L before the operation to respectively (13.1 ± 0.7) mmol/L and (12.3 ± 0.7) mmol/L at 4 and 8 weeks after the operation. Fasting insulin level increased from (28.0 ± 1.2) mU/L before the operation to respectively (62.8 ± 1.9) mU/L and (61.7 ± 1.4) mU/L at 4 and 8 weeks after the operation; and at 4 and 8 weeks after the operation postprandial insulin level was (77.4 ± 1.1) mU/L and (77.1 ± 1.0) mU/L. At 2 weeks from the operation, the fasting GLP-1 in group O increased from (10.7 ± 1.0) pmol/L to (13.5 ± 0.8) pmol/L, and respectively to (26.1 ± 0.9) pmol/L and (25.3 ± 1.2) pmol/L at 4 and 8 weeks after the operation. The differences in the above-mentioned items before and after the operation were all significant in group O (P < 0.05), and the differences in the items among group O and the other three groups (P < 0.05) were all significant as well. In group O, the apoptosis rate of pancreatic islet cell decreased to (5.9 ± 0.7)% at 4 weeks from the operation, and (6.3 ± 1.1)% at 8 weeks from the operation (P < 0.05). The expression of Bcl-2 protein in group O was 31.3 ± 1.5, 35.7 ± 1.0 and 35.8 ± 0.8 at 2, 4 and 8 weeks post operation, which was significantly higher in statistics than those of the same time point in the other three groups (P < 0.05). The expression of Bax protein in group O was 13.3 ± 0.9, 10.8 ± 0.9 and 10.9 ± 1.1 at 2, 4 and 8 weeks from the operation, which was significantly lower in statistics than those of the same time point in the other three groups (P < 0.05).

CONCLUSIONSGastric bypass surgery can significantly reduce the blood glucose level and promote the secretion of GLP-1, and therefore inhibit the apoptosis of the islet β cells in diabetic rats through the Bcl-2 pathway.

Animals ; Apoptosis ; Blood Glucose ; Diabetes Mellitus, Type 2 ; pathology ; surgery ; Disease Models, Animal ; Gastric Bypass ; Glucagon-Like Peptide 1 ; blood ; Insulin ; blood ; Islets of Langerhans ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the characteristics of B cell-activating factor (BAFF) secreted by peripheral blood monocyte-derived dendritic cell (MoDC) in chronic idiopathic thrombocytopenic purpura (cITP) and the function of MoDC on B cell proliferation.

METHODSTen cITP patients were studied dynamically before and after treatment. The BAFF levels in serum and the supernatant of LPS stimulated MoDC were tested with ELISA. The BAFF gene expression in LPS stimulated MoDC was tested with RQ-PCR, the B cell proliferation co-cultured with the supernatant of LPS stimulated MoDC for 5 days was tested with flow cytometry for CFSE and (3)H thymidine incorporation.

RESULTSThe BAFF level in serum (serum BAFF) \[(2461 ± 483) ng/L\], and supernatant of LPS stimulated MoDC (supernatant BAFF) \[(1113 ± 113) ng/L\] and BAFF mRNA in LPS stimulated MoDC (BAFF mRNA) (1.70 ± 0.23) before treatment were higher than that after treatment \[(621 ± 53) ng/L, (490 ± 49) ng/L and 0.37 ± 0.12\] and normal group \[(742 ± 77) ng/L, (582 ± 63) ng/L and 0.52 ± 0.08\]. There was a positive correlation among serum BAFF, supernatant BAFF and BAFF mRNA, and a negative correlation among serum BAFF, supernatant BAFF and BAFF mRNA and blood platelet count (BPC) in all ITP patients. The supernatant of LPS-stimulated MoDC from untreated patients enhanced B cell proliferation as compared with the supernatant of LPS-stimulated MoDC from treated patients and normal group.

CONCLUSIONBAFF might contribute to disease development in cITP. MoDC may directly increase B cell proliferation by secreting BAFF without T cell help, playing an important role in the antibody production in cITP.

B-Lymphocytes ; immunology ; Dendritic Cells ; immunology ; Humans ; Interleukin-4 ; Monocytes ; Purpura, Thrombocytopenic, Idiopathic ; immunology

AIMTo investigate protective effects of ginkgolide B (GB) in different administration modes on glutamate-induced neuronal damage.

METHODSEssential GB were obtained by supercritical CO2 fluid extraction. Glutamate excitotoxicity were examined in primary cultures from neonatal Wistar rat, by using of Trypan blue dye staining, testing the lactate dehydrogenase leakage from cultured neurons and terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) method. The protective effects of GB in different administration modes (pre-treatment and post-treatment) were adopted and compared with the NMDA receptor uncompetitive antagonist-MK-801 in acute-treatment.

RESULTSTreatment with GB in two administration modes both could increase ratio of surviving neuron, decrease LDH efflux and reduce ratio of neuron apoptosis in different degree, depended on dose in certain range. The protective effect of pre-treatment was superior to post-treatment, but inferior to MK-801.

CONCLUSIONGB can protect neurons against glutamate damage, and preventive using has more efficiency. The potential mechanism of its neural protection may be not only related to PAF receptor. If the predominant protection effect of GB in pretreatment is considered, precautionary intervention to high-risk population could have more value.

Animals ; Cells, Cultured ; Dizocilpine Maleate ; pharmacology ; Ginkgolides ; administration & dosage ; pharmacology ; Glutamic Acid ; adverse effects ; Hippocampus ; drug effects ; metabolism ; Lactones ; administration & dosage ; pharmacology ; Neurons ; drug effects ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo study the complete genome sequence of Japanese encephalitis virus (JEV) strain XJ69 isolated in ZheJiang province and explore its evolution.

METHODSOverlapping primers were designed according to the full-length genomes from GenBank. RT-PCR was used to amplify the fragments and RT-PCR products were cloned T vector, sequenced and analyzed.

RESULTSThe genome of strain XJ69 and XJP613 were 10 964 nucleotides in length with a single open reading frame encoding 3432 amino acids. Comparison of the complete genome sequences of different JEV isolates showed XJ69 and XJP613 were 83.5%-99.2% and 83.4%-99.4% nucleotide sequence homology among them respectively, which resulted in 94.8%-99.7% amino acid sequence homology. Phylogenetic analysis through PrM/C,E and full-length genome showed that the XJ69 and XJP613 strain belonged to genotype I.

CONCLUSIONThe nucleotitede sequence and deduced amino acid sequence of XJ69 and XJP613 strain were similar to that of those of genotype I of Japanese encephalitis virus. It belonged to genotype I and were close to the isolates SH17M-07.

Animals ; Cell Line ; China ; Cricetinae ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Encephalitis, Japanese ; virology ; Genome, Viral ; Humans ; Molecular Sequence Data ; Phylogeny