OBJECTIVETo investigate the dose and time-dependent effects of lipopolysaccharide (LPS) on cytoskeletal F-acitn and G-actin reorganizations by visualizing their distribution and measuring their contents in human umbilical vein endothelial cell line ECV-304.

METHODSF-actin was labeled with rhodamine-phalloidin and G-actin with deoxyribonuclease I (DNase I)conjugated with fluorescein isothiocyanate (FITC). Contents of cytoskeletal proteins were obtained by flow cytometry.

RESULTSF-actin was mainly distributed peripherally in endothelial cells under normal conditions. LPS stimulation caused the formation of stress fibers and filopodia. G-actin was normally seen in perinuclear and nuclear areas in control ECV-304 cells. Under LPS stimulation, G-actin dots appeared in the cytoplasmic region. The actin disorganization was accompanied by the time- and dose- dependent decrease in F-actin pool and increase in G-actin pool.

CONCLUSIONSLPS can induce characteristic morphological alterations of actin cytoskeleton and formation of intercellular gap in endothelial cells, accompanied by changes in F-actin and G-actin pools.

Actins ; drug effects ; Analysis of Variance ; Cells, Cultured ; Deoxyribonuclease I ; Dose-Response Relationship, Drug ; Endothelial Cells ; chemistry ; Escherichia coli ; Fluorescein-5-isothiocyanate ; Fluorescent Dyes ; Humans ; Lipopolysaccharides ; pharmacology ; Phalloidine ; Rhodamines ; Umbilical Veins ; cytology

OBJECTIVEThis research aimed to study and assess clinical application of the using of three Nickel-titanium (NiTi) rotary instruments, namely ProFile, ProTaper and Hero 642, in preparation of curved root canals.

METHODS80 teeth with curved root canals were instrumented by ProFile, ProTaper or Hero 642 rotary instruments using crown-down technique in test groups and by stainless steel K files in control group. All teeth were obturated with lateral condensation method. The efficiency of preparation and obturation was analyzed with radiographs before, during and after operation.

RESULTSThree NiTi rotary instruments could keep the curvature and flow of curved canals very well, and the continuously tapered root canal shape was achieved. There were no transportation, apical blockage, and ledge in all test groups. The operative time was shorter for ProTaper than that of ProFile and Hero 642. Endodontic flare-up seldom occurred in test groups. Two rotary instruments fractured in canals.

CONCLUSIONWith the NiTi rotary instruments, ProFile, ProTaper and Hero 642, curved root canals can be prepared effectively and quickly. Meanwhile, the problem of instrument separation should be prevented.

Dental Alloys ; Dental Pulp Cavity ; Humans ; Molar ; Nickel ; Root Canal Preparation ; Root Canal Therapy ; Stainless Steel ; Titanium

OBJECTIVETo evaluate clinical effect of the new ProTaper Nickel-titanium rotary instruments in preparation of curved root canals.

METHODS68 teeth with curved root canals were instrumented by ProTaper rotary instruments using crown-down technique, and by K files using step-back technique for control. All teeth were obturated with lateral condensation method. The efficiency of preparation and obturation was analyzed with radiographs before, during and after operation.

RESULTSNo transportation, apical blockage and ledge were found in test. The technique could keep the curvature and flow of curved canals. There were two ledge and more apical transportation (P < 0.01) in control than in test. The operative time was shorter and post treatment pain seldom occurred in ProTaper group.

CONCLUSIONThe ProTaper NiTi rotary instruments can prepare curved root canals effectively and safely. It is an efficient instrumentation method for curved canals and can be used widely.

Dental Instruments ; Humans ; Nickel ; Root Canal Preparation ; instrumentation ; Titanium

AIM(1) To investigate the mRNA expression of the key angiogenic growth factors in the grafts after transplantation. (2) To investigate the potential impact of danshen (Chinese traditional medicine) administration on grafts angiogenesis.

METHODSThe frozen-thawed ovarian tissue from aborted fetus were xenografted into the renal capsule of the nude mice, recovered 48 h, 7 d and 28 d after respectively. Either danshen or saline (as the control) was administered after transplantation.

RESULTSThe mRNA levels of VEGF showed a temporary raise in 48 h after transplantation, then decreased in one week, and no significant difference was fund between the control group and danshen group. Ang-2 was increased in 48 h after transplantation, when Danshen group was significantly higher than the control group (P < 0.05). The microvessel density significantly increased in all the tissues after transplantation. The control group peaked on day 7 after transplantation, while danshen group peaked in 48 h and kept correspondingly steady after that.

CONCLUSIONEarly angiogenesis began within 48 h after transplantation of the thawed human fetal ovarian tissue, and its microvessel density peaked within the first week after transplantation. Our results also suggested that the use of danshen injection in conjunction with transplantation could facilitate revascularization of the grafts.

Angiopoietin-2 ; genetics ; metabolism ; Animals ; Cryopreservation ; Drugs, Chinese Herbal ; pharmacology ; Female ; Fetal Tissue Transplantation ; methods ; Fetus ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Physiologic ; drug effects ; Ovarian Follicle ; cytology ; growth & development ; transplantation ; RNA, Messenger ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; Transplantation, Heterologous ; methods ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

BACKGROUNDEarly intensive insulin therapies in newly diagnosed type 2 diabetic patients may improve beta-cell function and yield prolonged glycemic remissions. This study was performed to evaluate the relationship between the glycemic remission and beta-cell function and assess the variables predictive of long-term near-normoglycemic remission.

METHODSEighty-four newly diagnosed type 2 diabetic patients were treated with 2-week continuous subcutaneous insulin infusion (CSII) and followed up longitudinally. Intravenous glucose tolerance tests (IVGTTs) were performed, and blood glucose, hemoglobin A1c (HbA1c) and insulin were measured at baseline, after CSII and at 2-year visit. The patients who maintained glycemic control for two years were defined as the remission group and those who relapsed before the 2-year visit were the non-remission group.

RESULTSThe duration to be diagnosed of the patients (from the time that patients began to have diabetic symptoms until diagnosis) in the remission group was shorter than that in the non-remission group (1.00 month vs 4.38 months, P = 0.040). The increase of the acute insulin response (AIR) was maintained after 2 years in the remission group compared with AIR measured immediately after intervention (413.05 pmol*L(-1)*min(-1) vs 408.99 pmol*L(-1)*min(-1), P = 0.820). While AIR in the non-remission group significantly declined (74.71 pmol*L(-1)*min(-1) vs 335.64 pmol*L(-1)*min(-1), P = 0.030). Cox model showed that a shorter duration to be diagnosed positively affected the duration of near-nomoglycemic remission with an odds ratio (OR) 1.019, P = 0.038, while fasting plasma glucose (FPG) and post-breakfast plasma glucose (PPG) after CSII were the risk factors (OR 1.397, P = 0.024 and OR 1.187, P = 0.035, respectively).

CONCLUSIONThe near-normoglycemic remission is closely associated with long-term maintenance of beta-cell function and occurs more commonly in patients with shorter duration to be diagnosed and better glycemic control during CSII.

Adult ; Aged ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; pathology ; Female ; Follow-Up Studies ; Humans ; Hyperglycemia ; pathology ; Hypoglycemic Agents ; therapeutic use ; Insulin ; therapeutic use ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Young Adult

OBJECTIVE: To investigate the features and diagnosis way of small round cell tumor of nasal sinus and bases of skull. METHOD: A retrospective analysis of 123 case with small round cell tumor of nasal sinus and bases of skull were carried out in our hospital in past ten year. Clinical, histological, radiological and immunohistochemical characters of these cases were studied. RESULT: All cases usually complained of nasal obstruction, headache, diplopia, nasal mucus with bleeding, vision or weight loss. Expansible or infiltrative lumps were found in nasal sinus or bases of skull in radiological examination. A lot of small round cells were found in these tumors in histological pathology. At least 5-6 cell, tissue or tumor markers were examined immunohistochemically in most of cases before the final diagnosis were made. In some cases over 20 markers were examined. Five cases were carried out transmission electron microscope examination, special features such as desmosome and myofilament were found. CONCLUSION Clinical symptom, physical signs and radiological finding can supply malignant evidences of these tumors. Histological examination can make certain that they are small round cell tumors, but final diagnosis is still hard to make only by these. Immunohistochemical examination of various markers can tell the original characters of the specimen tissues, it is the key for final diagnosis. Transmission electron microscope examination is another helpful way for diagnosis.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Leukemia, Lymphocytic, Chronic, B-Cell ; diagnosis ; pathology ; Male ; Middle Aged ; Paranasal Sinus Neoplasms ; diagnosis ; pathology ; Retrospective Studies ; Skull Base Neoplasms ; diagnosis ; pathology ; Young Adult

【Objective】To investigate the impact of long- term storage time on epigenetic modification of histone in human cleavage stage embryos.【Methods】According to the length of storage time，donated embryos after slow-freezing were divided into 3 groups ：6-year group ，9-year group ，and 12-year group ，while the control group consisted of donated fresh embryos. Immunocytochemistry was performed to compare the expression levels of HDAC1， H3K9ac， H3K4me3 ，and H3K9me3 among 4 groups. Transcription levels of HDAC1 ，SUV39H1 ，SETDB1 ，and KDM5A were analyzed through Single-Cell qRT-PCR.【Results】The relative abundances of HDAC1 and SUV39H1 mRNA showed no significant differences among 4 groups（P > 0.05）. SETDB1 exhibited a climbing pattern as storage time increased，but no significant difference was observed（P > 0.05）. There were no differences in H3K9 trimethylation and H3K9 methylation among 4 groups. However ，the expression level of KDM5A increased with the increasing storage time（P < 0.05）.【Conclusions】 Storage time did not affect the expression of deacetylase HDAC1，methylase SUV39H1 and SETDB1. H3K9ac/me3 and H3K4me3 also exhibited no significant difference as the storage time increases. However ，the increasing storage length might induce the elevating expression of demethylase KDM5A，which may be associated with inhibition of embryonic transcription.

OBJECTIVETo explore the expression and clinical significance of Hedgehog signaling transcription factor Gli1 in acute lymphoblastic leukemia (ALL) patients.

METHODSThe clinical specimens were obtained from 32 newly diagnosed and 6 relapsed ALL patients. Normal bone marrow cells from 15 healthy donors were used as controls. Real-time qPCR and Western blot were applied to detect Gli1 mRNA and protein expression in bone marrow mononuclear cells (BMMNC) of these samples respectively. The relation of Gli1 mRNA levels with clinical parameter was also evaluated.

RESULTSThe expression level of Gli1 mRNA in de novo and relapsed ALL patients was significantly higher than that in the normal controls (P < 0.05). There was no stalistically significant difference of the Gli1 mRNA expression between de novo and relapsed ALL cases (P > 0.05). In 24 de novo ALL patients with complete remission (CR) after induction chemotherapy, the levels of Gli1 mRNA were significantly reduced as compared with levels before treatment (P < 0.05). However, in 4 ALL patients without remission, no obvious difference of Gli1 mRNA levels were observed as compared with levels of Gli1 before treatment (P > 0.05). A positive correlation between the Gli1 mRNA expression level and white blood cell count (WBC) was found in the BMMNC of ALL patients (R = 0.725, P < 0.05). Similarly, Gli1 protein expression was significantly higher in the de novo and relapsed ALL cases compared with normal controls. The Gli1 protein level was down-regulated when the ALL patients was in CR.

CONCLUSIONThe expression of Gli1 mRNA and protein has been found to be high in de novo and relapsed ALL patients, and the change of Gli1 expression maybe relate to therapeutic efficacy and prognosis of ALL patients.

Bone Marrow Cells ; Humans ; Induction Chemotherapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prognosis ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Remission Induction ; Transcription Factors ; Zinc Finger Protein GLI1

OBJECTIVETo assess the accuracy and reliability of the nest-PCR-sequence specific primer(SSP) method in HLA-A site genotyping of single blastomeres retrieved from human pre-implantation embryos.

METHODSBy nest PCR on HLA-A exon 2, the success rate of first-round amplification was estimated for single blastomeres. Based on the first-round amplification, the HLA-A genotype of every single blastomeres was analyzed by commercially available PCR-SSP kits.

RESULTSThe amplification of HLA-A exon 2 were performed to 120 blasotmeres retrieved from in vitro fertilization(IVF) surplus embryos donated by 10 couples. The average success rate of family 1-5 and 6-10 was 78.2%(43/55) and 93.8%(61/65), respectively. And 86.7%(104/120) in total. Eighty blastomeres were further tested by nest-PCR-SSP, among which 11 blastomeres failed to HLA-A exon 2 amplification and then failed to genotyping while the other 69 blastomeres succeed in HLA-A exon 2 amplification and succeed in genotyping. Except for 6 blastomeres that were uncertain for allele lost because of parents' homozygosity, the left 63 blastomeres had accurate HLA genotyping. Among these 63 blastomeres, 59 blastomeres had genotypes confirmed from their parents(93.6%), 3 blastomeres lost one of parents' alleles(4.8%), and only one blastomere had two more than parents' alleles(1.6%).

CONCLUSIONThe above research results indicated that based on the successful first round amplification of single blastomeres, nest-PCR-SSP strategy offers a convenient and reliable option for HLA genotyping on single blastomeres, which is a key process in pre-selecting HLA-identical sibling for allogeneic cord blood cell transplantation.

Base Sequence ; Blastomeres ; metabolism ; DNA ; analysis ; DNA Fingerprinting ; methods ; DNA Mutational Analysis ; Female ; HLA Antigens ; analysis ; HLA-A Antigens ; analysis ; genetics ; Histocompatibility Antigens Class I ; analysis ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Single Person

Child ; Cord Blood Stem Cell Transplantation ; Hematopoietic Stem Cell Transplantation ; Humans ; Primary Myelofibrosis/therapy* ; Thiotepa ; Transplantation Conditioning