Objective To investigate the effects of X-ray dose on the expressions of microRNA-221 （miR-221） and phosphatase and a tensin homolog deleted from chromosome10 （PTEN） in human colorectal carcinoma （CRC） cells. Methods Human CRC-derived cell line， Caco2， was cultured conventionally. The cells were divided into five groups and exposed to different doses of X-ray （0， 2， 4， 6， and 8 Gy） respectively. The total RNA and protein of the Caco2 cells were extracted after irradiation， and the miR-221 and PTEN mRNA expressions were detected by real-time RT-PCR.Moreover， the protein alteration of PTEN in Caco2 cells was detected by Western-blot analysis. Results The radiation dose of X-ray significantly affected the expressions of miR-221 and PTEN protein in human Caco2 cells in a dose-depen-dent manner. Moreover， the miR-221 expression level was up-regulated gradually with the increase of irradiation dose， on the contrary， the PTEN protein expression level was down-regulated gradually （P＜0.01）. Conclusion The radiation dose can affect the miR-221 and PTEN protein expression pattern in CRC cells.
Objective To investigate the effects of matrine on proliferation, apoptosis and radiotherapy sensitivity of uveal melanoma cells. MethodsAn animal experiment study. In vitro experiment: MuM2B cells of human choroidal melanoma were randomly divided into control group and matrine 0.25, 0.50, 1.00, 2.00 g/L groups. The cell morphology was observed by transmission electron microscope. Cell proliferation was detected by thiazole blue colorimetry. The mRNA and relative expression levels of CyclinD D (CyclinD), B lymphoblastoma-2 (Bcl-2) and Bcl2-associated X protein (Bax) were detected by real-time polymerase chain reaction and Western blot. In vivo experiment: BALB/C mice were injected with MuM2B cell suspension subcutaneously on the back of forelimb to prepare transplanted tumor model. After successful modeling, they were randomly divided into blank group and matrine treatment group with different concentrations. Mice in blank group were injected with phosphate buffer subcutaneously. Mice in different matrine treatment groups were injected with 15, 25, 50, 100 mg/kg matrine subcutaneously, respectively, for 7 consecutive days. The tumor was weighed and its volume was measured after the last administration. Single factor analysis of variance was used to compare different groups. The t test was used for pairwise comparison between groups. ResultsIn the control group, the cell structure was normal, the distribution was uniform, and no or rare nuclear pyknosis was seen. With the increase of matrine dosage, the nuclear pyretosis increased gradually and cell morphology changed obviously. Compared with the control group, the cell survival rate in 0.50, 1.00 and 2.00 g/L groups gradually decreased with matrine concentration increasing and treatment time prolongating, the relative expression levels of CyclinD and Bcl-2 mRNA and protein gradually decreased, and the relative expression levels of Bax mRNA and protein gradually increased. Under the same radiation dose X-ray irradiation, the cell survival rate of 0.50, 1.00 and 2.00 g/L groups gradually decreased, and the differences were statistically significant (P＜0.05). Compared with blank group, the tumor weight and volume of mice in different doses of matrine group were significantly decreased, and the differences were statistically significant (P＜0.05). ConclusionMatrine can down-regulate the expression of CyclinD and Bcl-2, up-regulate the expression of Bax, promote the apoptosis of MuM2B human melanoma cells, inhibit cell proliferation, and enhance cell radiosensitivity.
Objective To investigate the radiation-sensitizing effects of Ku80 silencing by siRNA interference for A549 lung cancer cells. Methods The sequences of Ku80 siRNA and negative siRNA were chemically synthesized and transfected into A549 lung cancer cells by lipofectamine. RT-PCR and Western bolt analysis were used to determine Ku80 gene expression. The transfected cells in culture dishes were irradiated with X ray at doses of 2 Gy, 4 Gy, 6 Gy, 8 Gy, 10 Gy, respectively. Once all treatments were completed, the cells were processed with the colony formation assay. Results RT-PCR detection showed that Ku80 mRNA levels in A549 lung cancer cells were reduced after transfected with Ku80 siRNA at 24 h, 48 h and 72 h time points. Western blot analysis showed that Ku80 protein content decreased at 48 h and 72 h time points compared with the control group ( P lt; 0. 05 ) . Cloning formation assay indicated that radiosensitivity of A549 lung cancer cells was enhanced after transfected with Ku80 siRNA. Conclusion Ku80 siRNA can effectively inhibit Ku80 gene expression of A549 lung cancer cells, and therefore enhance its radiosensitivity.
Objective To summarize the current advancement of preoperative radiotherapy for rectal cancer. Methods Relevant literatures about current advancement of preoperative radiotherapy for rectal cancer published domesticly and abroad recently were collected and reviewed. Results The lower local recurrence rate and longer disease-free survival time were observed in preoperative radiotherapy， compared with postoperative radiotherapy for rectal cancer. The recurrence rate was higher in short-course radiotherapy， compared with conventionally radiotherapy for stageⅢrectal cancer， but there was no significant difference for stageⅡrectal cancer. The biology molecular such as p53， CEA， Cox-2， EGFR， and VEGF had shown to be radiosensitive. Conclusions The proposal of preoperative radiotherapy for rectal cancer， could be prone to conventionally radiotherapy. There are more screening targets for preoperative radiotherapy in extensive exploration of diverse radiosensitivity. Biology molecular， developed gene expression profiling， and gene chips for rectal cancer may contribute to the individualization treatment.
Objective To investigate the expression and clinical significance of soluble triggering receptor expression on myeloid cells-1（sTREM-1） in sepsis patients.Methods Serum concentrations of sTREM-1，procalcitonin（PCT），tumor necrosis factor alpha（TNF-α） and interleukin-10（IL-10） were measured by enzyme-linked immunosorbent assay，while high sensitivity C-reactive protein （hsCRP） level was detected by immunoturbidimetry in 68 patients with sepsis，40 patients with no-infective SIRS，and 20 normal individuals. The diagnostic and prognostic value of sTREM-1 and its comparison with PCT and hsCRP were analyzed. The sequential organ failure assessment （SOFA） score system was used to evaluate the severity of sepsis. The relationship between sTREM-1, PCT , hsCRP , SOFA score，TNF-α ,and IL-10 of the sepsis patients was analyzed，respectively. Results The differences in the serum concentrations of sTREM-1，PCT，hsCRP，IL-10 and IL-10/TNF-α ratio had statistical significance among three groups（Plt;0.05）. The differences in the serum concentration of TNF-α had no statistical significance among three groups （Pgt;0.05）. However，the serum levels of sTREM-1，PCT and hsCRP in the sepsis group were significantly higher than those in the SIRS group （Plt;0.05）. The receiver operating characteristic curve （ROC） analysis showed the area under the curve （AUC） for sTREM-1，PCT and hsCRP were 0.772 （95%CI 0.674-0.871），0.718 （95%CI 0.601-0.835） and 0.664 （95%CI 0.532-0.797），respectively. The serum levels of sTREM-1 and PCT in the non-survivors were significantly higher than the survivors in the sepsis group （Plt;0.01），but the differences in the serum concentration of hsCRP had no statistical significance between the non-survivors and the survivors in the sepsis group （Pgt;0.05）. There were significantly positive correlations between sTREM-1 and SOFA score，IL-10 or IL-10/TNF-α ratio（r value of 0.453，0.301，0.417，Plt;0.05），but no correlation between sTREM-1 and TNF-α（Pgt;0.05）. There was significantly positive correlation between PCT and SOFA score （r=0.436，Plt;0.05），while no relationship between hsCRP and SOFA score（Pgt;0.05）. Conclusions The serum level of sTREM-1 not only be valuable in the diagnosis of sepsis，but also may be used as a prognostic marker in sepsis，as it can reflect the severity of sepsis in certain degree. Furthermore，sTREM-1 or PCT may be superior to hsCRP in diagnosis，prognostic judgment and severity assessment of sepsis.
Objective To measure the level of circulating endothelial progenitor cells ( EPCs) in peripheral blood of patients with acute exacerbation of chronic obstructive pulmonary disease ( AECOPD) , and to explore the relationship between EPCs and severity markers of the disease and cardiovascular adverse outcome predictors.Methods Forty patients with COPD were recruited, including 27 at acute exacerbation phase and 13 with stable COPD from December 2010 to December 2011. Sixteen healthy nonsmokers were included as controls. Circulating EPCs were isolated by Ficoll density-gradient centrifugation and purified by Magnetic Activated Cell Sorting system. High-sensitivity C-reactive protein ( hsCRP) was estimated by using a latex immunoturbidimetric assay kit, and matrix metalloproteinase-9 ( MMP-9) was measured by enzymelinked immunosorbent assay ( ELISA) . Arterial blood gas analysis and echocardiograph were performed in the AECOPD patients. The correlations between circulating EPCs, lung function, and cardiovascular markers were investigated. Results Circulating EPCs were significantly lower in AECOPD and stable COPD patients compared with the healthy controls [ ( 5.1 ±2.6) ×103 /mL and ( 6.0 ±3.2) ×103 /mL vs. ( 9.0 ±4.3) × 103 /mL, Plt;0. 05] . EPCs had a weak correlation with hsCRP ( P = 0. 033) , but not with MMP-9. In the AECOPD patients, EPC counts were significantly inversely correlated with PASP ( pulmonary artery systolic pressure) and NT-proBNP ( amino-terminal pro-brain natriuretic peptide) levels, and positively with left ventricular ejection fraction. No correlations were found between EPCs and lung function, blood gas, hospital stays or smoking index. Conclusions Circulating EPCs were significantly lower in AECOPD patients compared with healthy controls, in which systemic inflammation might be involved. Decreased EPCs were correlated with cardiac dysfunction in patients with AECOPD, which may account for the increased cardiovascular risk in this population.
This paper is aimed to investigate the effect of rest-inserted loading on the mechanosensitivity of osteocytes. In the investigation, cultured MLO-Y4 osteocyte-like cells were strained on cyclic compressive force (CCF) by the self-made compressive loading device. Then we observed the effect of different rest periods-inserted loading (5 s, 15 s, 30 s, respectively) on the mechanosensitivity of osteocytes. We then determined the levels of secreted nitric oxide (NO) and prostaglandin E2 (PGE2) by Griess method and enzyme linked immunosorbent assay (ELISA), respectively. We then stained the cytoskeleton F-actin using immunofluorescence. We found that the expressions of NO and PGE2 in rest-inserted strained groups (>15 s) were significantly increased compared to those in the continuous strained group. And rest-inserted loading promoted the parallel alignment of stress fibers. It indicates that rest-inserted loading could promote the mechanosensitivity of osteocytes, and this might be related to the parallel alignment of stress fibers.
Objective To study the retinal light sensitivity of central visual field in normal children. Methods The QZS-Ⅱ automated perimetry was used to assess the visual field of centro-30deg;and centro-6deg; in normal or ametropic eyes in 60 eyes of 5～9 years old children. Results The mean sensitivity(MS)was not influenced by sex,age and laterality and ametropia of the eye.The normal type of dB distribution was obviously higher than the abnormal(P＜0.01).We set normal range as 30deg;MS＞19.3 dB, 6deg;MS＞22.5 dB.The abnormality of value or distribution didnprime;t appear in the same field. Conclusions In normal children,the dB distribution of visual field was mainly of the normal type.We suggest that in evaluating function of visual filed of the children,the dB distribution of centro-30deg;and centro-6deg;field and the value of MS should be included. (Chin J Ocul Fundus Dis, 1999, 15: 137-138)
OBJECTIVE:To investigate the value of psychophysical testing for the macular function in the diegnosis of diabetic retinopathy(DR). METHODS:To compare the testing results of macular light sensitivity and pattern visual evoked potential(P-VEP)of 30 eyes of 15 normal person with those of 82 eyes of 41 diabetic patients(27 eyes without DR,55 eyes with simple type DR ). RESULTS:The macular light sensitivity of diabetic patients is much lower than that of normal Control group(plt;0.05). In the diabetic group, 62.19% is abnormal in macular light sensitivity, 69.51% in P-VEP. CONCLUSION: Testing of macular light sensitivit y is helpful in finding of diabetic retinopathy and early deterioration of macular visual function in diabetics. （Chin J Ocul Fundus Dis,1996,12: 223-224）