Objective To explore the effect of low calorie metabolism on the survival of HeLa cells exposed to X-rays,and the influence of starvation on the antioxidative factors in the blood of rats after irradiation.Methods MTT method was used to evaluate the impact of different concentration glucose on the proliferation of HeLa cells.Colony formation assay was employed to detect the influence of glucose ( 1,5,10 and 25 mmol/L) on radiosensitivity of HeLa cells. Flow cytometry assay was used to analyze distribution of cell cycle and apoptosis.60 male SD rats were randomly divided into 6 groups with 10 rats each.Rats in every two groups were fed ad libitum,fasted for 24 h and fasted for 48 h,respectively.Rats in one group of each approach were respectively exposed to whole-body X-rays at 11 Gy. At 2 h after irradiation,all of rats were sacrificed and their venous blood was collected.Elisa kits were used to detect superoxide dismutase (SOD) and total antioxidant capacity (T-AOC).Results An increased viability was observed in HeLa cells treated with the glucose at low concentration ( ＜25 mmol/L),while HeLa cell growth was inhibited by glucose at doses of ＞25 mmol/L. Relevant to cells treated with 1 mmoL/L glucose,SERs (sensitive enhancement ratio) in cells exposed to 5,10 and 25 mmol/L glucose were 1.07,1.10 and 1.23,respectively. A reduction of G2/M and S arrests and apeptosis caused by 6 Gy X-ray irradiation were observed [(49.68 ±1.88)％ and (35.54±1.45)％ at G2/M phase,(16.88 ±1.22)％ and (10.23 ±1.65)％ atS phase,t=10.42,5.61,P＜0.05]and in the cells treated with 1 mmol/L glucose compared with cells treated with 25 mmol/L glucose [ ( 25.50 ± 0.95 ) ％ and (7.56 ± 1.07 ) ％,t =21.72,P ＜0.05 ].Without irradiation,calorie restriction exhibited a negligible influence on SOD and T-AOC in rats.However,after 11 Gy irradiation,compared with rats fed ad libitum,the levels of SOD and T-AOC were significantly increased in rats with calorie restriction ( t =40.32,42.78, P ＜ 0.05 ).Conclusions Calorie restriction has a certain radioprotective effect in vivo and in vitro.

Objective To study the genes which simultaneously participate in different carcinogenesis progression in lung adenocarcinoma for biomarkers identification. Methods 10 lung adenocarcinoma samples including pathologic stage Ⅰ,Ⅱ,Ⅲ were chosen for experiment and their matched normal tissues for control. After hybridization on 20 slides of microarray with 13824 genes, we analyze the expression profiles combined with pathologic stage and clinical prognosis by data mining. The genes differentially coexpressed in different stage and different prognosis samples were the target. Results 119 genes were identified. Among these targets, 26 genes have known to be related to lung cancer, 46 genes were unreported and 47 gene were new. Conclusions The 119 genes were very important during cancer occurrence and development and were the candidate biomarkers in lung adenocarcinoma.

The CrdS protein responding to the acidic adaptation was prokaryotic-expressed in our Laboratory to explore the regulatory mechanism in the acidic adaptation of Helicobacter pylori (H. pylori). The whole genomic DNA of H. pylori strain 26695 was abstracted and set as the template firstly. And then the hp1364 gene coding CrdS protein was amplified via the PCR technique. Then the clonal recombinant plasmid pUCm-T-hp1364 and the prokaryotic expression plasmid pQE30-hp1364 were built and identified by the methods of PCR, cutting with two enzymes and sequencing. After that, the plasmid pQE30-hp1364 was transferred into the E. coli XL1 blue and induced with IPTG. Using western blot and SDS-PAGE, it can be analyzed that the expressed recombinant protein existed mainly in the form of the inclusion bodies and its relative molecular mass was about 46 kDa. The successfully attained recombinant protein CrdS will provide the material to explore the regulatory mechanism in the acidic adaptation of H. pylori and the new way to resist the infection of H. pylori.
Bacterial Proteins ; biosynthesis ; genetics ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; metabolism ; Helicobacter pylori ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics

BACKGROUND: Cytogenetic evidence suggests that chromosomal alteration are not randomly occurring events and some malignancies are characterized by specific chromosome abnormalities, which provides cytogenetic basis for the expression of oncogenes.OBJECTIVE: To investigate the characteristics of chromosomal karyotype and marker chromosome of breast cancer cell lines Bcap-37and MCF-7 by means of G-banding chromosomal analysis.DESIGN: A controlled experiment with breast cancer cells as observation subjects.SETTING: Department of Medical Genetics, Peking University Health Science Center.MATERIALS: This study was carried out in the Department of Medical Genetics of Peking University Health Science Center from April 1991 to May 1992 using breast cancer cell lines MCF-7 and Bcap-37.METHODS: The chromosomes of human breast cancer cell lines Bcap-37and MCF-7 were obtained by growth synchronization induced by hypothermia and colchicines treatment. The cells at prometaphase or metaphase underwent G-banding chromosomal analysis. For each cell line, 50 to 60 mitotic figures were counted and 15 or 16 G-binding karyotypes were analyzed, including the mitotic figure at the level of about 320- and 500-band .MAIN OUTCOME MEASURES:Abnormality in chromosome number and structural aberration of the two breast cancer cell lines.RESULTS:The modal chromosomal number of Bcap-37 cell line was 63, of which 17 marker chromosomes had identifiable structure, as compared with 13 out of 56 chromosomes in modal number of MCF-7 cell line.CONCLUSION: Both of the two breast cancer cell lines have complex cytogenetic abnormality in the modal number and structure of the chromosomes, which might result in the rearrangement of DNA sequence of the cancer-related genes or DNA depletion, so as to play important roles in the pathogenesis and development of breast cancer.

Objective To explore the effect of ionizing radiation and cisplatin on the expression of Transducer of erbB2,1 ( TOB1 ) in human lung adenocareinoma SPCA-1 and LTEP-α-2 cell lines.Methods SPCA-1 and/or LTEP-α-2 cells were respectively irradiated with 2,4,6,8,and 10 Gy X-rays generated by a linear accelerator (with the source skin distance of 100 cm and dose rate of 200 cGy/min).The cell molecular samples were subtracted at 6,12,18,and 24 h after 6 Gy irradiation.For X-rays and cisplatin combination treatment,cells were divided into radiation group (6 Gy X-rays),cisplatin group (20 mol/L cisplatin),and combination group (6 Gy X-rays + 20 mol/L cisplatin).SPCA-1 and LTEP-α-2 cells without any treatment were used as control group.The relative levels of TOB1 mRNA and protein expression were detected by rigorously controlled semi-quantitative RT-PCR and Western blot analysis with quantitation of the mRNA/protein bands by densitometry.Results The expression level of TOB1 was significantly increased in both mRNA and protein level after radiation exposure (t =8.25-24.48,P ＜0.05).For time-dependent induction of TOB1,the expression was significantly increased 6 h after X-rays exposure (t =14.23-15.82,P ＜ 0.05 ).More significant increase of TOB1 expression was identified after the combination treatment of X-rays and cisplatin,compared to that of the radiation and/or cisplatin treatment alone (t =11.21-13.67,P ＜0.05).Conclusions lonizing radiation and cisplatin could upregulate the expression of TOB1 in lung cancer cells in both mRNA and protein levels.TOB1 may be a molecular target for ionizing radiation and cisplatin treatment in lung cancer and it may have potential implication in evaluating the curative efficiency of chemo-and radio-therapy.

Objective To build the method of computerized case classification,for the purpose of perfecting the diseases typing and classification,and supporting the management based on the quality and expenses of the disease.Methods 875 cases were defined into two types.Eight parameters were selected for non-surgery cases,namely the disease diagnosis,severity at admission,and age.For surgery cases,five parameters were defined,namely the disease diagnosis,operative quantity,and severity at admission.Then Fisher function was called into play to obtain the function descriminant equation,realizing computerized classification.Results The rate of matching was 86.2％ between computerized classification and manual classification.The high accuracy of function descriminant equation proves the satisfactory outcomes of the classification.Collusion The computerized classification is satisfactory in its outcomes,and therefore it can better quality of care and cost management of diseases in clinical practice.

Due to individual differences of the depth of anaesthesia (DOA) controlled objects, the drawbacks of monitoring index, the traditional PID controller of anesthesia depth could not meet the demands of nonlinear control. However, the adjustments of the rules of DOA fuzzy control often rely on personal experience and, therefore, it could not achieve the satisfactory control effects. The present research established a fuzzy closed-loop control system which takes the cerebral state index (CSI) value as a feedback controlled variable, and it also adopts the particle swarm optimization (PSO) to optimize the fuzzy control rule and membership functions between the change of CSI and propofol infusion rate. The system sets the CSI targets at 40 and 30 through the system simulation, and it also adds some Gaussian noise to imitate clinical disturbance. Experimental results indicated that this system could reach the set CSI point accurately, rapidly and stably, with no obvious perturbation in the presence of noise. The fuzzy controller based on CSI which has been optimized by PSO has better stability and robustness in the DOA closed loop control system.
Anesthesia ; methods ; Anesthesiology ; methods ; Anesthetics, Intravenous ; administration & dosage ; Feedback ; Fuzzy Logic ; Propofol ; administration & dosage

Objective To observe the effect of fasudil on cytoskeleton reconstruction in mouse podocytes induced by angiotensin Ⅱ (Ang Ⅱ),as well as to study the protective mechanism of fasudil in the pathological changes of podocytes.Methods Conditionally immortalized mouse podocytes were treated with Ang Ⅱ (10-7 mol/L).The podocytes were pre-incubated for 30 min or 60 min with various concentrations of fasudil (10-8,10-7,10-5 mol/L),then Ang Ⅱ (10-7 mol/L) were added and furtherincubated for 24 hours.The cytoskeleton distribution of podocyte as indicated by F-actin and synaptopodin was observed by fluorescence microscopy and Western blotting.At the same time,the activity of Rho signal pathway that mediates actin filament polymerization was analyzed by measuring the extent of Rho-associated coiled-coil-containing protein kinase 1 (ROCK-1) and myosin phosphatase-1 (MYPT1).Results Compared to the control group,AngⅡ disrupted the podocyte actin cytoskeleton and significantly decreased the expression of synaptopodin (P ＜ 0.05),while fasudil stabilized the actin filaments,and improved the synaptopodin expression (P ＜ 0.05).The expression enhancement of ROCK-1 and MYPT1 by Ang Ⅱ were reduced significantly by fasudil (P＜0.05).Conclusion The cytoskeleton reconstruction of podocytes can be induced by Ang Ⅱ and inhibited by fasudil,which suggests that the protective effect of fasudil may be partially contributed to the Rho/ROCK signaling pathway.

Objective To analyze the relationship between leukocyte function-associated antigen-1 (LFA-1) mRNA expression and corneal inflammation and discuss the inhibition reaction of nuclear factor-?B(NF-?B)inhibitor on LFA-1 mRNA expression.Methods The corneal suture or combined with subconjunctival injection models were constructed in BALB/C mice.The corneas of each group were excised 1,3,7 and 14 d after operation and LFA-1 mRNA expressions were investigated by reverse transcription polymerase chain reaction(RT-PCR).Results At 1,3 and 7 d after operation, the expressions of corneal LFA-1 mRNA in corneal suture combined with lipopolysaccharide (LPS) (corneal suture+LPs)subconjunctival injection group were higher than those in corneal suture group (P0.05).At 1 and 7 d after operation, compared with corneal suture+LPS group ,the expression of corneal LFA-1 mRNA increased,but it decreased at 3 and 14 d after operation in the group of corneal suture combined with LPS and pyrrolidine dithiocarbamate (PDTC) (P

Objective To study the effect of sanguinarine on the growth and radiosensitivity of ovarian cancer SK-OV-3 cells.Methods Cell growth was determined by MTT and clonogenic assay.Cell cycle analysis was performed by flow cytometry assay.The cell apoptosis was analyzed by Annexin V/PI assay.Results Sanguinarine inhibited SK-OV-3 cell growth in a dose-and time-dependent fashion and its IC50 values were 3.02 and 1.11 μmol/L at 24 and 48 h,respectively. Sanguinarine also significantly triggered a sub-G1 peak,an indicator of apoptosis,and caused a G0/G1 arrest.Furthermore,the cell apoptosis induced by X-irradiation was significantly increased at 6 Gy when the cells were pre-treated with sanguinarine,in which the early apoptotic population increased from 10.28％ to 43.28％ (t =19.41,P ＜0.01 ) and the late apoptotic population increased from 20.26％ to 30.80％ ( t =8.78,P ＜ 0.01 ).The multi-target click model was used to fit survival curves and the SER of sanguinarine treatment approached to 1.625 at the dose of D0. Conclusions Sanguinarine could inhibit SK-OV-3 cell growth by inducing apoptosis and cell cycle arrest and enhance cell radiosensitivity at low doses.