OBJECTIVETo study the effect of hydroxycamptothecin (HCPT) on apoptosis-inducing factor (AIF) expression and AIF translocation from mitochondria to the nucleus in human hepatocellular cancer cell SMMC-7721 during apoptosis.

METHODSAfter treatment with 80 mg/ml of HCPT, the cancer cells were stained with A0/EB to monitor their apoptosis. Their mitochondria was examined with electronmicroscopy and the AIF expression of the cells was tested by RT-PCR and Western blot. The translocation of AIF from mitochondria to the nucleus during apoptosis was analyzed by confocal microscopy.

RESULTSSMMC-7721 cells treated with HCPT showed chromatin condensation, nuclear fragmentation and mitochondria swelling. The mRNA and protein expression of AIF in treated and untreated SMMC-7721 cells were not significantly different. However, cells treated with 80 mg/ml HCPT for 6 h or 12 h showed massive translocation of AIF into the nuclei.

CONCLUSIONThese results show the important role the mitochondrial pathway of apoptosis plays in HCPT-induced tumor cell death, at least in SMMC-7721 cells.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; Apoptosis Inducing Factor ; biosynthesis ; genetics ; Camptothecin ; analogs & derivatives ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; Translocation, Genetic

Objective To understand the current situation of occupational exposure and occupational safety attitudes among nursing students in Kunming Medical University, and to find effective interventions to improve nurses' occupational safety,and to reduce occupational exposure.Methods 854 students in Kunming Medical University were investigated by a questionnaire survey, including Demographic characteristics, Occupational exposure conditions, Occupational safety attitude and Gathers, quit intention . The correlations between occupational safety attitudes and occupational exposure were analyzed with Simple Linear Correlation.Results There were 94.30%female respondents,and only 5.70%male respondents.The average age of respondents was 22.52±2.90 years. The Simple Linear Correlation analysis showed that correlation between Negative attitude and occupational exposure was positive correlated, the occupational exposure and Support from managers, and Optimistic were negative correlation ( <0.05) .Conclusions Occupational safety and attitude are related to occupational exposure. Nursing students should apply standardized operation to improve the professional quality and ability. Schools should strengthen the education of occupational protection for nursing students. Hospital management should regularly organize nurses to study occupational exposure protection knowledge,give full affirmation of the nurse's achievements in the work, and offer certain reward appropriately.

Objective To explore the changes of soluble tumor necrosis factor receptor type 1(sTNFR1)in children with acute virus encephalitis(VE)and its clinical significance.Methods The levels of tumor necrosis factor-?(TNF-?),sTNFR1 and neuron specific enolase(NSE)in cerebrospinal fluid were determined by enzyme-linked immunosorbent assay(ELISA)in 55 children who were admitted with VE,including 25 cases with severe VE(SVE)and 30 cases with mild VE(MVE).Fifteen cases without VE were as control group.Results The levels of TNF-?,sTNFR1 in cerebrospinal fluid in encephalitis children were significantly higher than that in control group(Pa

OBJECTIVESTo investigate the differentially expressed mitochondrial proteins in hydroxycamptothecin (HCPT)-treated SMMC-7721 cells by using quantitative proteome.

METHODSSMMC-7721 cell apoptosis was induced by HCPT and the mitochondria were isolated with a mitochondria isolation kit. Mitochondrial proteins labeled with a cleavable isotope-coded affinity tag were identified and quantified using two-dimensional liquid chromatography/tandem mass spectrometry.

RESULTSHighly purified mitochondria were obtained. Seventy-four mitochondrial proteins, which were statistically significantly altered (P less than 0.05) in HCPT-treated cells, were identified and analyzed. A total of 42 proteins were significantly down-regulated, and 32 were up-regulated in the cells that responded to apoptosis. The functions of these proteins were likely involved in cell energy metabolism, nucleic acid translation and transcription, cytoskeleton, etc.

CONCLUSIONOur results about the information of differentially expressed mitochondrial proteins in HCPT-treated cells and the control cells will help to understand the mechanism by which HCPT induces cell apoptosis. The integrated techniques we used in this study will be helpful to the investigation of subcellular quantitative proteomics.

Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; Humans ; Mitochondria ; metabolism ; Mitochondrial Proteins ; metabolism ; Proteome ; metabolism

OBJECTIVETo measure the quantities and apoptosis-related protein levels of B lymphocyte in the patients with immunorelated pancytopenia (IRP) and explore the action of B lymphocyte in the pathogenic mechanism of IRP.

METHODSQuantities of whole B lymphocytes and CD5+ B lymphocytes as well as the expressions of Fas and Bcl-2 in B lymphocytes in 35 patients with untreated IRP, 15 IRP patients in complete remission (CR), and 10 normal controls were assayed by flow cytometry.

RESULTSThe percentages of B lymphocyte and CD5+ B lymphocyte were significantly higher in untreated IRP patients than in CR IRP patients and normal controls (P < 0.05), and there was no significant difference between the latter two groups (P > 0.05). There was no significant difference of Fas expression in B lymphocyte among three groups (P > 0.05). The expression of Bcl-2 in B lymphocyte was significantly higher in untreated patients than in CR patients or normal controls (P < 0.01), and significantly higher in CR patients than in normal controls (P < 0.01). The apoptosis-related index was significantly lower in untreated patients than in CR patients or normal controls (P < 0.05), and significantly lower in CR patients than in normal controls (P < 0.05). The percentage of B lymphocyte was positively correlated with post-treated response time (r = 0.53, P < 0.01).

CONCLUSIONThe production of auto-antibodies in IRP patients probably has some relationship with the abnormal quantities of B lymphocyte and its subpopulations as well as with the inhibition of B lymphocyte apoptosis.

Adolescent ; Adult ; B-Lymphocytes ; cytology ; immunology ; Case-Control Studies ; Child ; Female ; Humans ; Lymphocyte Subsets ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology

OBJECTIVETo evaluate the application of nephron-sparing surgery (NSS) in the treatment of renal cell carcinoma (RCC).

METHODSThe clinical data of 221 RCC patients who received nephron-sparing surgery were retrospectively reviewed. Open surgery was performed for 203 cases and laparoscopic surgery for 18 cases. The renal arteries were blocked and ice fragments were used for 115 patients. Biopsy was performed on renal tissue adjacent the tumor for 136 patients. After operation 156 patients were treated with interferon and interleukin II. Among these 221 patients, 132 patients (59.7%) were followed for 3-56 months (mean: 28 months).

RESULTSThe mean distance from the edge of normal tissue to the tumor was 0.4 cm (0.2-1.0 cm). The mean volume of blood loss was 180 ml (50-1 000 ml). Pathological examination showed clear cell carcinoma (n = 195), chromophobe renal carcinoma (n = 12), papillary renal carcinoma (n = 7), cystic renal carcinoma (n = 6), and sarcomatoid clear cell carcinoma (n = 1). Biopsies in 136 patients showed negative results. Post-operative complications included leakage of urine (n = 1), hemorrhage or hematuria (n = 5), and infarction of blood vessel of lower limbs (n = 2). During the follow-up, no patients died for this reason, although tumor recurrence (n = 4, 1.8%) and metastasis (n = 3, 1.4%) were noted.

CONCLUSIONNSS is a safe and effective method to treat carefully selected RCC patients.

Adult ; Aged ; Carcinoma, Renal Cell ; surgery ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; surgery ; Male ; Middle Aged ; Nephrectomy ; methods ; Nephrons ; surgery ; Retrospective Studies ; Young Adult

Objective To investigate the prognostic sequelae in neontes with hypoxic-ischemic encephalopathy (HIE) and ven-tricnlar dilatation.Methods Seventy-six full term newborns infants with HIE were followed up at the age from 3 to 19 months after therapy. Twenty-five infants among them were followed up by telephone in the epidemic period of SARS.Results Among 76 infants of 88 newborn infants with HIE(84.6%), 73 infants were normal (96.1% ). 1 infant had cerebral palsy (1.3%), 2 infants died (2.6 %).Among 39 cases with mild HIE, none of them had cerebral sequelae; among moderate HIE. 1 infant had cerebral palsy (2.9%) 1 infant died (2. 9 %), interlenkin-4 among severe HIE 50 % died (P00.5 The poor outcome of HIE in those infants were related to intrauterine growth retardation,severe birth asphyxia;and inadequate treatment.Cranial ultra-sonography of 49 infants were done on follow-up,and 12 of them (24.5 % ) had ventricular dilatations, which appeared after birth with 6 infants. Others occurred on follow-up with 1 infant had cerobral palsy,all ventricular dilatations recovered to normal at 12- 19 months except the cerebral palsy.Conclusions The poor outcome of HIE depends on the infants with intranterine growth relarda-tion,severe birth asphyxia and inadequate treatment.The prognosis of transient ventrealar ddatation are good except cerebral palsy.J Appl Clin pediatr,2004,19(12) : 1045- 1047

OBJECTIVETo investigate the differentially expressed mitochondrial proteins in hydroxycamptothecin (HCPT)-treated SMMC-7721 cells by comparative proteomic analysis.

METHODSApoptosis of SMMC-7721 cells were induced by using HCPT and their mitochondria were isolated with a mitochondria isolation kit for cultured cells. Three different solubility protein fractions were extracted with ReadyPrep Sequential Extraction Kit and were separated by two-dimensional gel electrophoresis (2-DE). PDQuest software was used to differentiate mitochondrial proteins between control cells and HCPT-treated cells. Matrix assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS) was used to identify some of the different proteins.

RESULTSHighly purified mitochondria and high resolution 2-DE patterns of the proteins were obtained. Forty-four mitochondrial protein spots from the HCPT-treated cells showed different expressions compared to those of the control cells. Twenty of the different protein spots were analyzed by MALDI-TOF-MS.

CONCLUSIONDifferently expressed mitochondrial proteins in HCPT-treated cells and control cells were obtained in this study. This will be of help to understand the mechanism by which HCPT induces cell apoptosis.

Antineoplastic Agents, Phytogenic ; pharmacology ; Camptothecin ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; Humans ; Membrane Potentials ; drug effects ; Mitochondrial Proteins ; metabolism ; Proteomics

OBJECTIVESTo explore the category, quantity and clinical significance of autoantibodies on bone marrow hematopoietic cells in patients with immunorelated cytopenia and evaluate the sensitivity of direct antiglobulin reaction (Coombs test ) of bone marrow mononuclear cells (BMMNC).

METHODSThe category and the positive rate of autoantibodies on bone marrow hematopoietic stem cells, nucleated erythrocytes, granulocytes in 32 patients with uncertain immunorelated cytopenia were investigated by using BMMNC-Coombs test and double immunofluorescence flow cytometry.

RESULTSThe positive rate of autoantibodies on bone marrow hematopoietic cells tested by flow cytometry was 90.63% which was higher than that by BMMNC-Coombs test (50.0%) (p < 0.05). In 29 positive cases, IgG autoantibody accounted for 6.90%, IgM13.8%, IgG+IgA 3.4%, IgG+IgM 31.0%, and IgG+IgM+IgA 44.8%. Of the 29 Patients, 25 (86.2%) with IgG autoantibody, 26 (89.7%) with IgM and 14 (48.3%) with IgA. The patients with IgG autoantibody alone had the lowest hemoglobin levels, and those with IgM autoantibody might have intravascular hemolytic findings. The response time of patients with IgG and IgG+IgM was shorter than that of the other patients. 91.3% of the patients had autoantibodies on bone marrow hematopoietic stem cells and showed pancytopenia, and 50% of the patients had autoantibodies on nucleated erythrocytes and granulocytes. Eleven of 13 patients with negative BMMNC-Coombs tests had autoantibodies on bone marrow hematopoietic stem cells detected by FACS. There was no significant difference of the quantities of the three categories of autoantibodies of nucleated erythrocytes and stem cells. The quantities of IgA on granulocytes were lower than that of IgG and IgM. There was no significant difference between IgG and IgM on granulocytes. The quantity of IgA on hematopoietic stem cells was significantly higher than that on nucleated erythrocytes or granulocytes.

CONCLUSIONSThe sensitivity of double immunofluorescence flow cytometry assay was higher than that of BMMNC-Coombs test for detecting autoantibodies. In immunorelated cytopenia patients, the predominant autoantibody was IgM which could cause intravascular hemolysis, and the second one was IgG which could cause severe anemia. Most immunorelated cytopenia patients had autoantibodies on hematopoietic stem cells and showed pancytopenia. IgA was more easily seen on the hematopoietic stem cells.

Adolescent ; Adult ; Aged ; Autoantibodies ; classification ; metabolism ; Autoimmune Diseases ; immunology ; Bone Marrow Cells ; immunology ; Child ; Coombs Test ; Female ; Flow Cytometry ; Humans ; Immunoglobulin A ; metabolism ; Immunoglobulin G ; metabolism ; Immunoglobulin M ; metabolism ; Male ; Middle Aged ; Pancytopenia ; immunology

OBJECTIVETo ascertain an accurate approach to inserting the pedicle screw into C3-C7 segments of the cervical vertebra.

METHODSAnatomic morphology of lateral mass and pedicle, and their anatomic relationship with the adjacent tissue were observed on C3-C7 segments of 25 adult embalmed cadavers (50 sides).

RESULTS1) The inferior edge of the base of the posterior tubercle of the transverse process and the inferior edge of the pedicle were connected with each other on 25 adult embalmed cadavers (50 sides). The transverse section which passed through the median point between the superior edge and the inferior edge of the base of the posterior tubercle of the transverse process, and the transverse section which passed through the central axis between the superior edge and the inferior edge of the pedicle, were in the same horizontal plane. The superior and inferior position of placing the pedicle screw was determined by this transverse section, which passed through the median point between the superior and the inferior edge of the base of the posterior tubercle of the transverse process. 2) There was a directed internal-downwards "triangular sulcule" between the base of the posterior tubercle of the transverse process and the anterolateral edge of the inferior articular process. The anterior wall of the triangular sulcule was the base of the posterior tubercle of the transverse process, the posterior wall was the anterolateral edge of the inferior articular process, and the bottom of the sulcule was connected with the interior edge of the pedicle. The vertical length between the top of triangle and the planes of inferior edge of the pedicle was (2.78+/-1.71) mm. The inferior edge of the cervical pedicle could be detected using a blunt probe along the "triangular sulcule" between the base of the posterior tubercle of the transverse process and the anterolateral edge of the inferior articular process in surgical operation. 3) The lateral fovea of the articular process was observed on all lateral masses (50 sides). The internal and external position of the entrance point could depend on anatomic landmarks: the lateral edge of the lateral fovea of the articular process. The horizontal length between the lateral fovea of the articular process and the entrance point was (3.14+/-1.45) mm. 4) The diameter of pedicle screw, about (2.78+/-1.71) mm, was the transverse diameter of the cancellous bone of the greatest narrow part of the cervical pedicle.

CONCLUSIONSThe median point between the superior edge and the inferior edge of the base of the posterior tubercle of the transverse process, the lateral fovea of the articular process, and the triangular sulcule between the base of the posterior tubercle of the transverse process and the anterolateral edge of inferior articular process, are easy to be exposed and identified in surgical operation. The pedicle screw can be precisely inserted through this method.

Adult ; Bone Screws ; Cervical Vertebrae ; anatomy & histology ; surgery ; Humans