0.05). Conclusions The expression of XAF1 decreases in both malignant and benign colo-rectal tumors and is significantly lower in colorectal carcinomas than in adenomas/polyps. Thus, XAF1 may have the potential role in differentiating colorectal cancer from benign tumors.

AIM: To investigate the potential role of 14-3-3 tau in trophoblast cells on invasiveness. METHODS: 14-3-3 tau expression was detected in first-trimester villi, deciduas and human trophoblastic cell line (BeWo) by immunohistochemistry. Small interference RNA (siRNA) targeting 14-3-3 tau was transfected into BeWo cells. The effects of down-regulated 14-3-3 tau on invasion of human trophoblasts cell line BeWo were examined by matrigel invasion assay, and the transcription, translation of E-cadherin and snail were estimated by RT-PCR or Western blotting. U0126 was used to detect the extracellular-signal related kinase 1/2 (ERK1/2) function on down-regulation of 14-3-3 tau induced cell invasion. RESULTS: 14-3-3 tau was detected in the invasive trophoblastic cells in the first trimester villi and that invaded to the deciduas. BeWo cells also expressed 14-3-3 tau. Down-regulation of 14-3-3 tau increased the invasive cell-number of BeWo, as well as the expression of snail, and inhibited E-cadherin. U0126 inhibited the enhanced invasiveness in these cells induced by the down-regulation of 14-3-3 tau. CONCLUSION: 14-3-3 tau may regulate the invasiveness of human trophoblastic cells through ERK1/2 signaling pathway.

Objective To evaluate the relationship between pathogenesis of preeclampsia (PE) and the ultrastructure change of the endoplasmic reticulum in trophocyte, mRNA and protein expression levels of endoplasmic reticulum molecular chaperone glucose-regulated protein 78 (GRP78), glucose-regulated protein 94 (GRP94), endoplasmic reticulum apoptosis factor cysteine protease protein 12 (caspase-12).Methods Sixty-five pregnant women who were hospitalized in the First Affiliated Hospital of Nanjing Medical University from July 2008 to January 2010, were selected as the subject. Thirty pregnancy women diagnosed with PE were divided into PE group and 35 normal pregnant women were used as control group.Electron Microscopy was used to measure ultrastructure change of the endoplasmic reticulum in placenta trophocyte. Reverse transcription(RT) PCR and western blot were used to investigute the expression levels of GRP78, GRP94, caspase-12 mRNA and protein in placenta. Results (1) In control group the volume of endoplasmic reticulum does not increase; no swelling and no expansion of endoplasmic reticulum was found.In PE group the edema number of endoplasmic reticulum was reduced; the volume of endoplasmic reticulum increased; expansion and vacuolation of cavity and degranulation of the endoplasmic reticulum was observed significantly. (2) The mRNA and protein expression levels of GRP78 in placenta of PE group (2.59 ± 0. 09 and 0. 81 ±0. 31) were significantly higher than those in placenta of control group (1. 16 ±0. 07 and 0. 40 ± 0. 10, P <0. 01). (3) The mRNA and protein expression levels of GRP94 in placenta of PE group (1.31 ± 0. 91 and 0. 55 ±0. 24) were significantly higher than those in placenta of control group (0. 63 ±0. 57 and 0. 22 ±0. 09, P < 0. 01). (4) The mRNA and protein expression levels of caspase-12 in placenta of PE group (4. 03 ± 0. 65 and 1.56 ± 0. 17) were significantly higher than those in placenta of control group (1.85 ± 0. 85 and 0. 91 ± 0. 69, P < 0. 01). Conclusion The obvious expansion of endoplasmic reticulum in trophocyte and the increased expression levels of GRP78, GRP94 and caspase-12 indicate that endoplasmic reticulum stress-mediated apoptosis may be involved in the pathophysiological processes of PE.

Objective: To study the effect of miR-200c on the migration and proliferation abilities of breast cancer MDA-MB-231 and BT-549 cells, and to clarify the mechanism of miR-200c in inhibiting the epithelial-mesenchymal transiton (EMT) of triple negative breast cancer. Methods: The human triple negative breast cancer cell lines (MDA-MB-231 and BT-549) were chosen in this study. The cells were transiently transfected with miR-200c mimics and Lipo2000 (experimental group), miR-200c negative control and Lipo2000 (negative control group), and Lipo2000 alone (reagent control group); at the same time, blank control group was set up. The expression levels of vimentin and (3-catenin mRNA and protein were detected by RT-PCR and Western blotting method. The proliferation rates and migration abilities of MDA-MB-231 cells and BT-549 cells after transfection of miR-200c were analyzed by CCK8 assay and wound healing assay. Results: The RT-PCR and Western blotting showed that the expression levels of vimentin and (3-catenin mRNA and proteins in experimental group were decreased, and the differences were statistically significant compared with blank control group, negative control group and reagent control group (P<0. 05). The CCK8 results showed that the proliferation rates of the cells in experimental group were lower than those in negative control group and reagent control group (P<0. 05). The wound healing assay results showed that the recovery rate of scratch width in experimental group was lower than those in negative control group and reagent control group (P<0. 05). Conclusion: miR-200c might inhibit EMT in triple negative breast cancer by regulating the expressions of (3-catenin and vimentin mRNA and proteins in MDA-MB-231 and BT-549 cells and decreasing the abilities of migration and proliferation of triple negative breast cancer cells.

With numerous advancements in novel biochemical techniques, our knowledge of the role of RNAs in the regulation of cellular physiology and pathology has grown significantly over the past several decades. Nevertheless, detailed information regarding RNA processing, trafficking, and localization in living cells has been lacking due to technical limitations in imaging single RNA transcripts in living cells with high spatial and temporal resolution. In this review, we discuss tech-niques that have shown great promise for single RNA imaging, followed by highlights in our recent work in the development of molecular beacons (MBs), a class of nanoscale oligonucleotide-probes, for detecting individual RNA transcripts in living cells. With further refinement of MB design and development of more sophisticated fluorescence microscopy techniques, we envision that MB-based approaches could promote new discoveries of RNA functions and activities.

Objective To detect the immune effect of FbaAmAb2 against the surface protein of group A Straptococcus (GAS),and explore the pathogenesis and therapy of GAS infections.Methods By subclonal and bacterial ELISA,the positive hybridoma cells were screened that can produce better titers of FbaAmAb2 against GAS-surface FbaA protein,and were injected into the peritoneal cavities of BALB/c mice to produce ascites.The collected ascites were performed to dilute,as follows,original ascite,1:2,1:4,1:8,and 1:16 to test tube agglutination.Based on the results,we selected appropriate dilution to passively immunize mice,and then challenged the mice with GAS,evaluating FbaAmAb2 neutralizing ability with GAS in mice by the survival rate of the immunized mice.Whether FbaAmAb2 could inhibit the binding of factor H to GAS was confirmed by the invasive inhibition assay.Results The IgG titer of bacteria solution ELISA is 1:160 and the titer of tube agglutination is 1∶8.The protect rates of FbaAmAb2 on preventing mice with GAS infections are as follows:66.67％ in original ascite and 1:2 diluted groups,and 50％ in 1:4 diluted group.Mice in each experimental group were evoked significantly protective immune responses compared with the PBS control by SPSS analysis.FbaAmAb2 can competitively inhibit factor H binding to the surface proteins FbaA of GAS,which decreased the entry of GAS into the cytoplasm of human epithelial cells through the binding of factor H.Conclusion FbaAmAb2 is promising to be used in emergent prevention or the clinical therapy for GAS infection and it is promising starting points for pharmacologic targeting and further development of new therapeutic agents for GAS.

Intracranial aneurysms are local abnormal bulging of intracranial arterial wall caused by various reasons. Since the International Subarachnoid Aneurysm Trial (ISAT) in 2002 confirmed the safety and effectiveness of endovascular therapy, interventional materials and treatment concepts have been continuously innovated, and endovascular therapy has become the first-line treatment of intracranial aneurysms. This article reviews the interventional materials and their progress in the endovascular treatment of intracranial aneurysms.

OBJECTIVEThe conversion of arginine into citrulline, termed citrullination, has important consequences for the structure and function of proteins. The present study aimed to identify novel citrullinated proteins in 10 tumor cell lines by 2-D Western blotting (2-D WB).

METHODSTwo identical two-dimensional electrophoresis (2-DE) gels were prepared using extracts from ten cultured human tumor cell lines: ECA(esophageal cancer cells), HEPG2 (hepatocellular carcinoma cells), SKOV3 (ovarian cancer cells), MCF-7 (breast cancer cells), H292 (lung mucoepidermoid carcinoma cells), HeLa (cervical cancer cells), Lovo (colon cancer cells), OS-RC (renal cell carcinoma cells), PANC-1 (pancreatic cancer cells), and SGC (gastric cancer cells). The expression profiles on one 2-DE gels were trans-blotted to PVDF membranes, and the blots were then probed with an anti-citrulline antibody. By comparing the 2-DE profile with the parallel 2-D WB profile at a global level, protein spots with immuno-signals were collected from the second 2-DE gel and identified using mass spectrometry. Immunoprecipitation was used to verify the expression and citrullination of the targeted proteins in the tumor cell lines.

RESULTS2-D WB and mass spectrometry identified citrullinated ENO1 (α-enolase), HSP60 (heat shock protein 60), KRT8 (keratin 8), TUBB (tubulin beta), TCRβ (T cell receptor β chain), VIME (vimentin) and PDI in these cell lines. Immunoprecipitation analyses verified the expression and citrullination of ENO1, HSP60, KRT8, and TUBB in the total protein lysates of the tumor cell lines.

CONCLUSIONThe citrullination of proteins ENO1, HSP60, KRT8, and TUBB suggests a new mechanism in the tumorigenic process.

Blotting, Western ; Cell Line, Tumor ; Citrulline ; metabolism ; Female ; Humans ; Immunoprecipitation ; Mass Spectrometry ; Phosphopyruvate Hydratase ; Vimentin

Objective To investigate the correlation between CT parameters of erector spinae muscle(ESM)and pulmonary function in elderly patients with chronic obstructive pulmonary disease(COPD),and to analyze its predictive value for the prognosis of patients.Methods A total of 120 COPD patients were included as the case group(including 60 cases in stable stage and 60 cases in acute exacerbation stage),and 60 smokers were selected as the control group.The differences of ESM CT parameters and pulmonary function parameters in each group were compared.According to the prognosis of COPD,patients were divided into good prognosis group(n=106)and poor prognosis group(n=14),and the predictive efficacy of ESM CT parameters on the prognosis of COPD patients was analyzed.Results The pulmonary function parameters,ESM cross sectional area(CSA)(ESMCSA)and ESM local volume in the case group were significantly lower than those in the control group(P<0.05).ESMCSA and ESM local volume were positively correlated with inspiratory capacity(IC),vital capacity(VC),forced vital capacity(FVC)and forced expiratory volume in one second(FEV1)(P<0.001).The average muscle density of ESM was positively correlated with IC,VC and FVC(P<0.05),but not with FEV1.The area under the curve(AUC)of ESMCSA and ESM local volume in predicting poor prognosis of COPD patients was 0.769[95%confidence interval(CI)0.661-0.876]and 0.827(95%CI 0.734-0.919),respectively.Conclusion There is a certain correlation between the CT parameters of ESM and the pulmonary function parameters of COPD patients,among which the ESMCSA and the ESM local volume have high predictive efficacy for the prognosis of COPD patients.

Objective:To explore the application of C-reactive protein(CRP) to prealbumin (PA) ratio(CRP/PA) for diagnosis and prognosis evaluation of sepsis.Methods:By a retrospective study, a total of 95 sepsis patients (sepsis group) and 100 local infection patients(non-sepsis group) treated in Dongying People′s Hospital from September 2021 to September 2022 were enrolled. Sepsis patients were divided into survival group(57 cases) and death group (38 cases) according to the 28-day outcome. The clinical data were collected and CRP/PA was calculated. Multivariate Logistic regression and Cox regression were used to analyze the relationship between various indicators and the occurrence and prognosis of sepsis, and receiver operating characteristic (ROC) curve was drawn to evaluate the diagnostic and prognostic value of CRP/PA for sepsis. Kaplan-Meier survival analysis was used to evaluate the prognostic value of different CRP/PA ratios for patients with sepsis.Results:The systolic blood pressure, diastolic blood pressure, prealbumin were lower and heart rate, respiratory rate, CRP, CRP/PA, procalcitonin were higher in the sepsis group compared to the non-sepsis group: (117.27 ± 11.65) mmHg (1 mmHg = 0.133 kPa) vs. (123.26 ± 10.71) mmHg, (69.42 ± 8.58) mmHg vs. (75.44 ± 6.53) mmHg, (174.09 ± 24.77) g/L vs. (207.13 ± 34.31) g/L, (97.87 ± 12.73) bpm vs. (86.90 ± 10.19) bpm, 22.0(20.00, 25.00) times/min vs. 21.00(19.00, 23.00) times/min, (93.96 ± 19.64) mg/L vs. (77.56 ± 22.54) mg/L, 0.54(0.44, 0.65) vs. 0.37(0.28, 0.46), 3.35(2.16, 4.17) μg/L vs. 1.52(0.81, 2.16) μg/L, there were statistical differences ( P<0.05). Multivariate Logistic regression analysis showed that CRP/PA and procalcitonin were risk factors for sepsis ( P<0.05). The results of ROC curve showed that the area under the curve (AUC) of CRP/PA in diagnosis of sepsis was 0.821, the specificity and sensitivity was 76.0% and 93.7%, respectively. The diastolic blood pressure, prealbumin, neutrophil were higher and the heart rate, respiratory rate, CRP, CRP/PA, lymphocytes, procalcitonin were lower in the survival group compared to the death group: (71.76 ± 8.86) mmHg vs. (67.86 ± 8.10) mmHg, (181.46 ± 24.35) g/L vs. (163.05 ± 21.28) g/L, (63.46 ± 9.88) × 10 9/L vs.(57.13 ± 8.64) × 10 9/L, (95.68 ± 13.48) times/min vs. (101.16 ± 10.88) times/min, 22.00(19.50, 24.00) times/min vs. 24.00(20.00, 28.00) times/min, (88.09 ± 19.35) mg/L vs. (102.76 ± 16.75) mg/L, 0.46(0.41, 0.58) vs. 0.63(0.55, 0.72), 21.00(16.00, 30.00) ×10 9/L vs. 29.50(18.00, 37.30) ×10 9/L, 2.94(2.10, 3.97) μg/L vs. 3.82(2.21, 4.77) μg/L, there were statistical differences ( P<0.05). Multivariate Cox regression analysis showed that CRP/PA and procalcitonin were independent risk factors for the prognosis of sepsis ( P<0.05). The AUC of CRP/PA in predicting the prognosis of sepsis was 0.827, the specificity and sensitivity was 92.1% and 63.8%, respectively. Grouped by the cut-off of CRP/PA (0.48), the 28-day mortality rate of patients in the CRP/PA>0.48 was significantly higher than that of patients in the CRP/PA≤0.48, there was statistical difference ( P<0.01). Conclusions:CRP/PA ratio can be used as an index for diagnosis and prognosis evaluation of sepsis.