Objective　To evaluate the effectiveness of intratracheal tube suctioning by respiratory mechanics measurement.Methods　The indexes of respiratory mechanics,including respiratory rate,expiratory tidal volume,% leak around the tube,dynamic respiratory compliance and mean airway resistance were measured just before and 20 minutes after intraendotracheal tube suction in mechanical ventilated children with respiratory failure.Results　Fifty-two measurements was carried out in 11 patie nts.The mean values of respiratory risistance was (116.73±27.12)cmH2O/L.s before suctioning and (93.38±26.64)cmH2O/L.s after suctioning,with significant differrence(P＜0.01)；The mean values of % leak around the tacheal tube dereased from(18.12±4.12)% before suctioning to (8.71±3.76)% after suctioning (P＜0.05),The mean values of expiratory tidal valume was markedly increased from(7.31±2.12)ml/kg before suctioning to(5.72±1.2)ml/kg after suctioning(P＜0.01),The total respiraory rate markedly decreased after suctioning.Conclusion　The respiratory mechanics measurement is very useful to evaluate the airway patient.The removal of secretions is crucial in respiratory managemet.

Adult ; Aged ; Animals ; Antineoplastic Agents ; therapeutic use ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Leeches ; Lymphatic Metastasis ; Male ; Materia Medica ; Middle Aged ; Nasopharyngeal Neoplasms ; drug therapy ; pathology ; radiotherapy ; Phytotherapy

A method for the determination of Fe, Co, Mn and Ni in synthetic diamonds by inductively coupled plasma atomic emission spectrometry ( ICP-AES) was proposed. The synthetic diamond sample was decomposed completely, while the sample was burned in air at 1000 ℃ for 10 h, and then a mixed acid of H2 SO4 , aqua regia and HClO4 was used for the dissolving the residue of the sample. In this method, the limits of detection of Fe, Co, Mn and Ni were 0. 0147, 0. 0018, 0. 0006 and 0. 0027 mg/L, respectively. Under the optimum condition, Fe, Co, Mn and Ni in synthetic diamond sample were determined. The values of RSDs (n=7) were less than 0. 5%. The recoveries of added standard were 94. 0%-105. 0%.

A method for the simultaneous determination of oxygen and nitrogen in synthetic diamonds by inert gas high temperature extraction-impulse heating method was proposed. The sample weight, the selection of analysis power and the calibration curves of oxygen and nitrogen were discussed. Oxygen and nitrogen in analytical samples are determined. Values of RSDs (n=7) for oxygen and nitrogen were less than 4. 5% and 4. 0% respectively. The analytical results of oxygen and nitrogen obtained by the proposed method were in good agreement with those by inert gas fusion-impulse heating method.

Objective To study the expressions of vascular endothelial growth factor (VEGF) and Ki-67 antigen and their correlation in human brain astrocytorna. Methods Immunohistochemistry with SP method was employed to detect the expressions of VEGF and Ki-67 in 60 human brain astrocytoma tissue (grade Ⅰ-Ⅱ 28 cases, grade Ⅲ 20 cases, grade Ⅳ 12 cases) and 30 normal tissues adjacent to the tumors. Results VEGF and Ki-67 were not expressed in the normal brain tissues adjacent to the astrocytoma, but their expressions were detected in all the astroeytoma tissues of different grades (P<0.05). High-grade astrocytomas showed significantly higher VEGF and Ki-67 expression than low-grade astrocytomas (P<0.05), and a positive correlation was noted between VEGF and Ki-67 expressions (r=0.310, P<0.05). Conclusion VEGF and Ki-67 expressions can provide important evidence for evaluating the malignancy and clinicopathologieal grading of human astrocytoma.

OBJECTIVETo introduce the experience in the treatment of lower eyelid pouches orbital rim.

METHODSAn incision was made along the margin of lower eyelid and dissection was performed under the orbicularis muscle to expose the orbital septum and periosteum of lower orbital rim. The fat released from orbital septum was transposed just below the lower orbital rim and fixed on the periosteum. If lacrimal groove deformity was not corrected completely, the musculocutaneous flap, which may be excised beside the incision, was kept to correct the deformities further with only the muscle portion.

RESULTS72 cases with lower eyelid pouches complicated with lacrimal groove deformities were treated with transposition of orbital fat and orbicularis muscular flaps. Satisfactory results were achieved in all the patients after a follow-up period of 3-6 months.

CONCLUSIONIt is an effective and feasible technique to correct lacrimal groove deformities with transposition of orbital fat and orbicularis muscular flaps.

Adipose Tissue ; transplantation ; Aged ; Blepharoplasty ; methods ; Eyelids ; surgery ; Humans ; Orbit ; Periosteum ; surgery

BACKGROUND AND OBJECTIVEAdenovirus vectors were widely used in gene therapy for tumors. We used adenovirus vector to transfer small interfering RNA (siRNA) against vascular epithelium growth factor A (VEGF-A) molecules to mouse lung adenoma LA795 cells and used low dose of chemotherapeutic drugs to further elevate the infection efficiency of adenovirus vector in and therapeutic effect of RNAi on tumor cells.

METHODSLA795 cells were infected by Ad/EGFP and treated with different dosages of gemcitabin, epirubicin, cisplatin, or 5-fluorouracil (5-FU). Cells were observed under fluorescence microscope continuously using green fluorescent protein (GFP) as the reporter gene. The percentage of GFP-positive cells and fluorescent intensity were tested by flow cytometry to determine optimum concentrations of drugs. Ad/siVEGF-A containing VEGF-A siRNA was constructed. Real-time PCR and ELISA were applied to measure the expression level of VEGF-A after LA795 cells were infected by Ad/siVEGF-A and treated with 5-FU. The combination of Ad/siVEGF-A and 5-FU was also applied in treating subcutaneous tumor in mice.

RESULTSLow dose of 5-FU elevated the Ad/EGFP infection in LA795 cells significantly, and also enhanced the effect of Ad/siVEGF-A in down-regulating VEGF-A mRNA and protein levels in tumor cells. When used in tumor in vivo, the combination strategy repressed tumor growth effectively.

CONCLUSIONLow dose of 5-FU can enhance the capability of adenovirus infecting tumor cells and promote the efficiency of gene therapy by adenovirus.

Adenocarcinoma ; metabolism ; pathology ; Adenoviridae ; genetics ; Animals ; Antimetabolites, Antineoplastic ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Fluorouracil ; administration & dosage ; pharmacology ; Genetic Therapy ; Genetic Vectors ; Lung Neoplasms ; metabolism ; pathology ; Mice ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics ; physiology

OBJECTIVETo evaluate the relationship of chimerism status of cell subsets with engraftment, occurrence of acute graft versus host disease (aGVHD), graft rejection and disease relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSChimerism status in peripheral blood (PB) and bone marrow (BM) of 65 patients received allo-HSCT were monitored at regular intervals post-transplant. Fluorescence-activated cell sorter (FACS) was used to sort CD3(+)T lymphocytes in 65 cases, CD3(-)CD56(+)CD16(+)NK cells in 52 cases, CD15(+) granulocytes in 32 cases and CD19(+)B lymphocytes in 20 cases post transplants. The chimerism status of different lineage cells was analyzed by polymerase chain reaction amplification of short tandem repeats (PCR-STR).

RESULTSOn day +7, NK-cells donor chimerism (DC 55.5%) was higher than other cell subsets. T lymphocyte was the latest one to reach complete donor chimerism (CDC) with a median on day +21. Patients whose T lymphocytes donor chimerism was more than 70% on day +7 and more than 95% on day +14 had a high risk for acute aGVHD. In all cases except those with ALL, the decreased DC of T lymphocytes were observed before molecular or hematological relapse occurred.

CONCLUSIONSerial and quantitative T cell chimerism analysis provides a reliable and rapid screening method for the early detection of engraftment, graft rejection, disease relapse and occurrence of aGVHD, therefore, is a prognostic tool to identify patients at high risk of aGVHD and disease relapse following allo-HSCT.

Adolescent ; Adult ; Child ; Chimerism ; Female ; Graft Rejection ; immunology ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Recurrence ; T-Lymphocytes ; immunology ; Young Adult

OBJECTIVEBy inhibiting AML1 -ETO fusion gene expression in Kasumi-1 cells with RNAi, to investigate the changes in cell proliferation and cell cycle.

METHODSThe small interference RNAs (siRNAs) specifically targeting the AML1 -ETO fusion gene were synthesized in vitro and transfected into Kasumi-1 cells by electroporation, the non-specific siRNAs transfected cells were taken as control. EGFP plasmid was transfected into Kasumi-1 cell and the transfection efficiency was detected by FCM. Inhibitory effect of siRNAs were detected by real-time RT-PCR and Western blots. Cell proliferation was measured by CCK-8 assay. DNA content was detected by PI assay.

RESULTSThe transfection efficiency was 44.5%. The AML1 -ETO specific siRNAs inhibited AML1 -ETO expression at both mRNA and protein levels. The cell proliferation rate in siRNAs treated group was lower than that in control group 72 h after transfection [(47.90 +/- 0.02)% vs (66.90 +/- 0.08)% , P < 0.05]. The cell cycle was blocked at G1 phase 72 h after siRNAs treatment, the cell proportion in G1 phase being 38.3% and 31.6% in control group, while in G2/M phase being 1.8% and 2.4% respectively.

CONCLUSIONSThe synthesized siRNAs can inhibit AML1 -ETO fusion gene expression. AML1 -ETO specific siRNA induced the decline of AML1 -ETO fusion protein in Kasumi-1 cell, and then caused the cell cycle blocked in G1 stage and eventually inhibited the cell proliferation.

Cell Cycle ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; Humans ; Leukemia ; genetics ; metabolism ; pathology ; Oncogene Proteins, Fusion ; genetics ; metabolism ; RNA Interference ; RUNX1 Translocation Partner 1 Protein ; Transfection

OBJECTIVETo investigate the effects and related mechanisms of hepatitis B virus X (HBx) protein on cell cycle and growth in hepatocellular carcinoma.

METHODSA human hepatocyte HepG2 cell line stably expressing a green fluorescent protein (GFP)-tagged HBx (HepG2/GFP-HBx cells) was used for the experiment, and HepG2 parental and HepG2/GFP cells was used as the controls. Effect of HBx on cell growth was evaluated by the MTT cell proliferation assay and on cell cycle progression by flow cytometry analysis of cells with or without treatment with 5-aza-2'-deoxycytidine (5-Aza-CdR; 5 pmol/L). Effect of HBx expression on promoter methylation status of the p16INK4A tumor-suppressor gene was detected by methylation-specific polymerase chain reaction and on p16 protein level was analyzed with western blotting.

RESULTSThe HepG2/GFP-HBx cells showed significantly higher cell proliferation at 72 hrs of culture (3.225+/-0.038 A490) than either control (HepG2: 2.012+/-0.022 A490, t = -46.86, P less than 0.001; HepG2/GFP: 2.038+/-0.029 A490, t = 42.51, P less than 0.001). The HepG2/GFP-HBx cells also showed significantly lower proportion of cells in the G0/G1 phase (16.45%+/-0.45%) than either control (HepG2: 44.81%+/-1.36%, t = -34.202, P less than 0.001; HepG2/GFP: 42.76%+/-1.58%, t = -28.88, P less than 0.001). However, 5-Aza-CdR treatment did lead to a significant amount of HepG2/GFP-HBx cells being arrested in the G0/G1 phase (33.25%+/-0.79%, t = 31.85, P less than 0.001). The p16INK4A promoter was methylated in the HepG2/GFP-HBx cells, and became demethylation after treatment with 5-Aza-CdR. However, no methylation of p16INK4A promoter was observed in both HepG2 and HepG2/GFP cells. The p16 protein level was significantly lower in the HepG2/GFP-HBx (vs. HepG2 and HepG2/GFP cells) and this level increased after treatment with 5-Aza-CdR.

CONCLUSIONHBx protein promotes hepatocellular carcinoma cell cycle progression and growth by shortening the G0/G1 phase, and the underlying mechanism may involve inducing p16INK4A promoter methylation and downregulating p16 protein expression.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Hep G2 Cells ; Hepatitis B virus ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Promoter Regions, Genetic ; Trans-Activators ; pharmacology