Objective:To compare the sealing ability and marginal adaptation of three obturation techniques when used in root canal obsturation of c-shaped canals of mandibular second molars. Methods:Twenty-one extracted mandibular second molars with C-shaped canals were randomly divided into three groups. After instrumentation, the canals were filled using three different techniques: ObturaⅡ(group A), cold lateral condensation (group B) and Thermafil (group C). Then, gaps,obturated lateral canals and reticular apical triangles on the X-ray were counted. After dyeing for 7 days in the ink, dyeing lines were measured to reflect the microleakage. The percentages of gutta-percha, sealer and gaps on the root canal surface were determined by analyzing the images of 3 sections per tooth. Results:The reticular apical triangles were most frequently observed in group C(P

Apoptosis ; Ceramides ; biosynthesis ; Sphingomyelins ; metabolism

Objective Idiopathic pulmonary fibrosis ( IPF) is a chronic inflammatory disease with unknown etiology and is lack of effective therapy. The aim of this study is to explore the function of Ca2+ and PI3K/AKT/mTOR pathway in the pathogenesis of IPF, and the impact of 1,25?( OH) 2 D3 on Ca2+ and PI3K/AKT/mTOR pathway in type Ⅱalveolar epithelial cells of rat with IPF. Methods 150 male SD rats were randomly divided into 2 groups: prevention group ( control groupⅠ, model groupⅠ, medication groupⅠ) and treatment group ( control groupⅡ, model groupⅡ, medication groupⅡ) . The tracheal exposure surgery was operated in control groupⅠ/Ⅱ, and then 200μL sterile physiological saline was administered by intraperitoneal injection of each rats 2 days and 14 days after surgery, separately. Bleomycin(BLM)(5 mg/kg) was in?jected into the trachea of model groupⅠ/Ⅱ, and then vitamin D3 solvent(0.1%ethanol and 99.9%glycol propylene, 1μL/g) was ad?ministered by intraperitoneal injection 2 days and 14 days after surger?y, separately. Bleomycin( BLM) ( 5 mg/kg) was injected into the tra?chea of medication groupⅠ/Ⅱ, and then 1,25?( OH) 2 D3( 2μg/kg) was administered by intraperitoneal injection 2 days and 14 days after surgery, separately. IPF model was built by injecting Bleomycin into the trachea of rats, 1,25?(OH)2D3(2μg/kg) was used to prevent and treat IPF by intraperitoneal injection in medication group. The hydroxyproline content of lung tissue in each group was measured, type Ⅱalveolar epithelial cells were separated from lung tissue and labeled with Fluo?3AM, then concentration of Ca2+ was detected by Laser scanning confocal microscope. The mRNA levels of PI3K, AKT and mTOR in the typeⅡalveolar epithelial cells were tested by RT?PCR. Results Compared with control groupⅠ/Ⅱ at each time point, hydroxyproline content of lung tissue, Ca2+ concentration and expression of PI3K, AKT and mTOR in typeⅡalveolar epithelial cells in model groupⅠ/Ⅱand medication groupⅠ/Ⅱwere sig?nificantly raised( P<0.05 or P<0.01) , but these were significantly reduced in medication groupⅠ/Ⅱcompared with model groupⅠ/Ⅱ( P<0.05 or P<0.01) . Correlation analysis showed that there is significant positive correlation between Ca2+ concentration and mRNA expression levels of PI3K, AKT and mTOR in model groupⅠ/Ⅱ(r=0.5988, r=0.6230, r=0.6603,P<0.01)and medication groupⅠ/Ⅱ( r=0.701 2, r=0.632 3,r=0.740 3,P<0.01) . Conclusion The PI3K/AKT/mTOR pathway plays an important role in devel?opment of IPF. 1,25?( OH) 2 D3 is able to reduce Ca2+concentration in typeⅡalveolar epithelial cells and inhibit the PI3K/AKT/mTOR pathway, and then inhibit the development of IPF.

Objective:To systematically assess the efficacy and safety of bevacizumab (BEV) plus chemotherapeutic agents as first-line therapy for metastatic colorectal cancer (mCRC). Methods:We retrieved randomized controlled clinical trials (RCTs) of BEV plus chemotherapeutic agents as first-line therapy for mCRC from the databases of PubMed (1966 to August 2009), Embase (1974 to August 2009), Cochrane Library (Issue 3, 2009), China Journal Full Text Database (CJFD, 1994 to August 2009), Chinese Bio-medical Literature Database(CBM, 1978 to August 2009) and Chinese Scientific Journals Full Text Database (CSJD, 1989 to August 2009). Then we evaluated the methodological quality of included studies. Meta-analysis was performed using RevMan 5.0 software developed by the Cochrane Collaboration. Results:Only 6 clinical studies were selected and 2 646 eligible patients were included in the systematic review. Meta-analysis showed that BEV plus chemotherapy increased the overall response rate (complete response+partial response) compared with chemotherapy alone. The relative risk was 1.27 (95%CI: 1.00-1.61, P=0.05), and the median survival time and progression-free survival (PFS) were longer. In terms of safety, there was a significant difference in the frequency of grade 3/4 adverse events, grade 3/4 hypertension, adverse events-induced study discontinuation and gastrointestinal perforation between the two groups. The relative risk was 1.12 (95%CI: 1.07-1.61), 4.51 (95%CI: 2.81-7.23), 1.37 (95%CI: 1.16-1.63)and 4.32(95% CI:1.24-15.05), respectively. There was no statistical difference between the two groups in the incidence of grade 3/4 bleeding, 60-day all-cause mortality, grade 3/4 proteinuria, grade 3/4 diarrhea, grade 3/4 leukopenia and pulmonary embolism. The relative risk was 1.50(95%CI: 0.87-2.57), 0.71(95%CI: 0.45-1.11), 2.26(95%CI: 0.69-7.33), 1.18(95%CI: 0.99-1.41), 1.17 (95%CI: 0.97-1.42)and 0.84(95%CI: 0.46-1.53), respectively. Conclusion:BEV plus chemotherapeutic agents as first-line the-rapy increases the response rate and prolongs PFS and median survival time of mCRC patients, but results in a higher incidence of any grade 3/4 adverse event, grade 3/4 hypertension and gastrointestinal perforation.

Objective:The recombinant human retinal pigment epithelium-derived factor(PEDF)protein to be obtained and the angiogenesis of the rPEDF to be identified.Methods: PEDF gene gene was amplified by PCR and cloned into pET32a,rPEDF protein was expressed in E.coli BL21 and confirmed by SDS-PAGE and Western blot.The rPEDF was purified by Ni-NTA on denature condition.The concentration of the rPEDF was determined by Bradford method.The angiogenesis of the rPEDF was determined by chick chorioallantoic membrane(CAM) method.Results: The expression plasmid pET32a-PEDF was constructed successfully.The rPEDF was expressed with stable efficiency in E.coli BL21.The results of the CAM experiment showed that the rPEDF had notable angiogenesis effect in the concentration 0.4、0.04 ng/ml,but had no effect in 4 ng/ml.Conclusion:The PEDF gene was cloned and expressed efficiently,the angiogenesis of the rPEDF to be identified and the activity was worked in certain range.The results can facilitate studying its function and spreading its application.

Objective To investigate the role of calreticulin (CRT) in the pathogenesis of rheumatoid arthritis (RA) by analyzing its expression in the sera,synovial fluid and synovial membrane in patients with RA.Methods Levels of CRT in the sera from patients with RA,osteoarthritis (OA),systemic lupus erythematosus (SLE),other autoimmune diseases and health control (HC) were detected by enzyme linked immunosorbent assay (ELISA) and Western bloting.CRT levels in synovial fluid from RA and OA patients were measured by ELISA.Pathological methods were employed to analyze the expression and localization of CRT in synovial membrane.ANVOA,SNK-q test were used for statistical analysis.Results CRT was found to be significantly up-regulated in sera from RA [(4.8±2.4) ng/ml] than that from in OA [(3.6±0.9)ng/ml],SLE [(4.0±1.5) ng/ml],other autoimmune diseases [(3.9±0.8) ng/ml] and HC [(3.7±0.6) ng/ml].Only the monomer form of CRT in the serum could be detected.Pathological results showed that in RA synovium,CRT was mainly expressed in the lining and sublining layers,endothelial cells and perivascular areas； while in OA,only few CRT staining was seen in perivascular areas and the synovial lines.Conclusion In RA,increased levels of CRT are detected in the sera.In addition,high expression of CRT is observed in synovium and its distribution are different from OA.Our results suggest that CRT may be involved in RA joint inflammation and pannus formation.CRT may become a potential serological marker in the diagnosis of RA.

Objective:To explore the function of IL-9 and PU.1 on genesis and development of pulmonary fibrosis,and the effect of active vitamin D3[1,25(OH)2VD3] on the expression levels of this two factors during the pathogenesis of fibrosis.Methods: 90 male SD rats were randomly divided into model group,treatment group,control group (n=30).Bleomycin(5 mg/kg) was injected into the trachea of rats to establish the model of pulmonary fibrosis in the model group and treatment group,while the control group was injected with isopyknic sterile saline.The treatment group,the model group and the control group were injected intraperitoneally with active vitamin D3,solvent of vitamin D3 (propylene glycol) and sterile saline on the 2nd day after surgery respectively.All injections were carried out once every other day.10 rats were euthanized at 14th,21st and 28th day in each group in turn.After obtaining lung tissues from experimental rats,the pathological change of lung was compared by hematoxylin-eosin staining.The difference of collagen fiber and hydroxyproline content were compared by the Masson staining and basic-hydrolysis method respectively.The mRNA and protein expression of IL-9 and PU.1 in lung tissue were detected by Real-time PCR and immunohistochemical technology respectively.The expression of IL-9 in serum was detected by ELISA.Results: Fibrosis appeared in lungs of experimental rats treated with bleomycin after 14 days,and more and more aggravated with time.At three time points,the hydroxyproline content in model group and treatment group were significantly higher than that of control group,and the treatment group was significantly lower than the model group.At three time points,the expression of IL-9 and PU.1 in model group and treatment group were risen gradually,and obviously higher than that in control group.On the 14th and 21st day,the expression of two factors in treatment group was significantly lower than model group;on the 28th day,there was no statistically significant difference between treatment group and model group(P>0.05).In model group and treatment group,the expression of two factors on 21st day was significantly higher than that on 14th day;there was no statistically significant difference between the 28th day and the 21st day.Conclusion: IL-9 and PU.1 may play a profibrotic role at early stage of pulmonary fibrosis induced by bleomycin.The active vitamin D3 may lower the expression level of PU.1,and then reduce the secretion of IL-9,thus may play an inhibiting effect on genesis and development of pulmonary fibrosis in rats.

Point Dingchuan (Ex-B1) pertains to extraordinary points and often produces a good therapeutic effect on respiratory system diseases. Ancient Chinese medical books do not record point Dingchuan. Modern Chinese medical textbooks describe the location and efficacy of and indications for this point but do not state their provenances. In order to further popularize the clinical application of point Dingchuan, this article verifies the origin and development of the name and location of point Dingchuan so as to provide a certain reference for clinically correct selection and use of this point. The results show that point Dingchuan followed the course of development from ashi point to new point and then to extraordinary point. The development of point Dingchuan to the same acupoint name and location as now international use went mainly through four stages, that is, similar name and different location, similar name and location, same name and different location, and same name and location.

Objective To study improvement of neural function by stereotaxic transplantation of adipose-derived stem cells (ADSC) into lateral cerebral ventricle after intracerebral hemorrhage in rats and its mechanism. Methods ADSC were cultured and proliferated in vitro, which had been marked with Brdu for 48 h before transplantation. The rat caudate nucleus hemorrhage (ICH) models were divided into 2 groups. ADSC were stereotaxically transplanted into the right lateral ventricles in ADSC group, and equal volume of saline was transplanted into control group. The score of neurological behavior were evaluated at modeling and 1, 3, 7, 14, 28 days after transplantation respectively.Double-staining immunofluorescence technique was used to detect Brdu-positive cells and the differentiation of neurons and astrocytes. In accordance with the instructions of TUNEL kit, cell apoptosis, and the expression of VEGF and angiogenesis were assayed. Results In vitro ADSC expressed undergo osteogenic and adipogenic differentiation. Compared with the control group, ADSC group had better motor function at 3, 7, and 14 days (P＜0. 05). Double-staining immunofluorescence showed mostly grafted Brdu-reactive ADSC had migrated to the hematoma zone, and some survivedand expressed Neun of Gfap. TUNEL analysis revealed that, 3 days after transplantation, the number of apoptotic cells in ADSC group was significantly less than in the control group (P＜0. 05). Three days after transplantation, VEGF expression levels in ADSC group were significantly higher than in the control group (P＜0. 05). Conclusion ADSC stereotaxially transplanted into the lateral ventricle can survive and differentiate into neuron-like cells. ADSC transplantation may reduce apoptosis and secret VEGF to promote the angiogenesis, and improve neural functional in intracerebral hemorrhage rats.

At present, acute myeloid leukemia (AML) accounts for about 15%-20% of childhood acute leukemia. Although overall survival rate is increasing with the help of risk stratification, stratification of chemotherapy, and supportive treatment, conventional pharmacotherapy still has a limited clinical effect and certain limitations in improving remission rate in previously untreated patients and reducing recurrence after remission. With the development of precision medicine, the mechanisms of targeted therapy, including abnormal activation of AML-related signaling pathways and epigenetic modification, have been found in recent years. Molecular-targeted drugs can therefore act on specific receptors and target genes to improve clinical effect and the prognosis of AML patients.
Child ; Epigenesis, Genetic ; Humans ; Immunotherapy ; Leukemia, Myeloid, Acute ; drug therapy ; mortality ; Molecular Targeted Therapy