A robust, selective and highly sensitive chemiluminescent （CL） platform for protein assay was presented in this paper. This novel CL approach utilized rolling circle amplification （RCA） as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier. Typically, the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence. This aptamer-primer sequence was then employed as the primer of RCA. Based on this design, a number of the biotinylated probes and streptavidin-horseradish peroxidase （SA-HRP） were captured on the plate, and the CL signal was amplified. In summary, our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized, and devoid of light source. Therefore, this new technique .will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases, by taking advantages of high sensitivity and selectivity.

A robust,selective and highly sensitive chemiluminescent (CL) platform for protein assay was presented in this paper.This novel CL approach utilized rolling circle amplification (RCA) as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier.Typically,the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence.This aptamer-primer sequence was then employed as the primer of RCA.Based on this design,a number of the biotinylated probes and streptavidin-horseradish peroxidase (SA-HRP) were captured on the plate,and the CL signal was amplified.In summary,our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized,and devoid of light source.Therefore,this new technique will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases,by taking advantages of high sensitivity and selectivity.

Objective: To observe the efficacy of mild moxibustion combined with arthroscopic meniscal repair in the treatment of meniscal injury and to explore its action mechanism. Methods: Ninety-eight patients with meniscal injury were divided into a surgery group and a moxibustion plus surgery group by the random number table method, with 49 cases in each group. Both groups received arthroscopic meniscal repair, and the moxibustion plus surgery group was treated with add-on mild moxibustion. The Lysholm score, visual analog scale (VAS) score, and mobility of the affected knee were measured before and after treatment, and transforming growth factor (TGF)-β1 and platelet-derived growth factor (PDGF) levels were measured in the fluid of the affected knee joint. The healing of the meniscus was recorded at a follow-up visit 3 months after the surgery. Results: After treatment, the Lysholm score increased, the VAS score decreased in both groups, and the intra-group differences in both groups were statistically significant (P<0.05). The Lysholm score was higher in the moxibustion plus surgery group than in the surgery group, and the VAS score was lower in the moxibustion plus surgery group than in the surgery group. The differences between groups in both scores were statistically significant (P<0.05). The mobility of the affected knee joint increased in both groups (P<0.05), and it was greater in the moxibustion plus surgery group than in the surgery group (P<0.05). When compared with pretreatment, the levels of TGF-β1 and PDGF in the fluid of the knee joint increased in both groups (P<0.05), and the levels of TGF-β1 and PDGF in the fluid of the affected knee joint were higher in the moxibustion plus surgery group than in the surgery group (P<0.05). The healing of the meniscus in the moxibustion plus surgery group was significantly better than that in the surgery group at the follow-up visit 3 months after the surgery (P<0.05). Conclusion: The effect of mild moxibustion combined with arthroscopic meniscal repair is better than the surgery alone in improving knee function, relieving pain, increasing mobility of the affected knee, and promoting meniscal healing, which may be related to the up-regulation of TGF-β1 and PDGF levels in the fluid of knee joint.

Objective To investigate the predictive value of early prognosis of the 40 Hz auditory steady-state response (40 Hz ASSR) in patients with coma following cardiopulmonary resuscitation (CPR). Methods Thirty patients with coma following CPR admitted in the Neurological Intensive Care Unit (NICU) were examined with the 40 Hz ASSR and shortlatency somatosensory evoked potential (SLSEP), and both were graded. Using transferred out of NICU as the short-term outcome end point, the patients with coma following CPR were divided into a survival group (n =21) and a death group (n =9; including brain death). The correlation between the 40 Hz ASSR and SLSEP grading and prognosis was analyzed. Results The grades of the 40 Hz ASSR (r = 0. 722, P = 0.000) or SLSEP (r = 0. 430, P = 0.018) was significantly correlated with the short-term prognosis. The sensitivity, specificity and accuracy of the 40 Hz ASSR for predicting the short-term prognosis were 77. 8%, 100% and 93.3%, respectively; and those of SLSEP were 88. 9%, 61. 9% and 70. 0%, respectively. Conclusions The 40 Hz ASSR has a certain prognostic value in patients with coma following CPR. The higher the grade of the 40 Hz ASSR is, the greater the likelihood of the recent death.

Objective To investigate the predictive value of prognosis of the 95% spectral edge frequency (SEF95) and total power (TP) in quantitative electroencephalography (qEEG) in patients with disturbance of consciousness. Methods The patients with disturbance of consciousness admitted in the neurointensive care unit (NICU) in Nanfang Hospital from January 2008 to June 2010 were included. Glasgow Coma Scale (GCS) scores were performed on admission and EEG monitoring was performed simultaneously. The patients were divided into either a survival group or a death group according to the survival status of the patients at the time of leaving NICU. The age, sex, hypertension, diabetes, GCS scores, SEF95, and TP were compared between the two groups. A multivariate logistic regression analysis was performed for the above factors. The prognostic indicators were analyzed with the receiver operating characteristic (ROC) curves and the of qEEG predictive ability of death in patients with disturbance of consciousness were determined. Results A total of 109 patients with supratentorial lesions were enrolled in the study, 79 of them were in the survival group and 30 of them were in the death group. The GCS scores (5 ±3vs. 9 ±3, P =0. 000) and SEF95 (7. 0 ±4.0 vs. 10. 0 ±4. 0, P = 0. 002) in the death group were significantly lower than those in the survival group. Multivariate logistic regression analysis showed that GCS scores (odds ratio 0. 100, 95% confidence interval 0. 029-0. 353) and SEF95 (odds ratio 0. 853, 95% confidence interval 0. 740-0. 983) were the independent predictors of recent prognosis. ROC curve analysis showed that the lower the GCS scores and SEF95 were, the greater the likelihood of death in patients. When SEF95 was ＜7. 75, the sensitivity to determine the death was 60. 0%, the specificity was 72. 2%, the positive predictive value was 82. 6%, and the negative predictive value was 45. 0%; when the GCS score was ＜8, the sensitivity to determine the death was 83. 3%, the specificity was 73. 4%,the positive predictive value was 82. 6%, and the negative predictive value was 45. 0%. Conclusions SEF95 helps determine the prognosis of patients with disturbance of consciousness, and it is expected to become an important means of bedside assessment of prognosis in patients with disturbance of consciousness in NICU.

BACKGROUND: Overexpression of phosphorylated Tau protein is a factor of dementia, and scholars abroad find that APP17 peptide may have effect on it.OBJECTIVE: To observe changes of phosphorylated Tau protein Ser202/Thr205 of mice with diabetes mellitus (DM) after injection of APP17 peptide.DESIGN: Randomized control study.SETTING: Department of Pathology, Capital University of Medical Sciences; Department of Brain Aging, Xuanwu Hospital, Capital University of Medical Sciences.MATERIALS: The experiment was carried out in the Pathological Department of Capital University of Medical Sciences and Brain Aging Department of Beijing Xuanwu Hospital. A total of 18 male Kunming mice of 8 weeks old and weighing 28-32 g were randomly divided into control group, DM group and APP17 peptide group with 6 in each group.METHODS: DM models were induced by streptozotocin (STZ) through selectively destroying β-islet cells; meanwhile, APP17 peptide was intraperitoneally injected into mice. Four weeks later, brain tissue underwentimmunohistochemical staining with AT-8 (Ser202/Thr205, a special monoclonal antibody).MAIN OUTCOME MEASURES: ① Morphological observation; ② AT-8 distribution; ③ quantitative analysis of immunohistochemical staining.RESULTS: Positive AT-8 cells in DM group were distributed in retrosplenial cortex, hippocampus, thalamus, hypothalamus, etc.; however, those incontrol and APP17 peptide groups were only distributed in retrosplenial cortex and hippocampus, and poorly stained.CONCLUSION: Positive AT-8 cells may be widely distributed in neurons of brains of DM mice; however, APP17 peptide may normalize the expression of positive AT-8 cells.

This study was undertaken to observe the biochemical changes in striatum of Parkinson's disease (PD) rat model by modified proton magnetic resonance spectroscopy. 12 SD rats were divided into model (n=7) and control (n=5) groups. At 3 weeks after the injection of 6-hydroxydopamine into right striatum, 1H-MRS on the striatum was taken by modified proton magnetic resonance spectroscopy, and then tyrosine hydroxylase (TH) immunostatining was used to visualize the changes of the neurons in substantia nigra and neurites in striatum. The results showed that TH positive neurons and neurites in the substantia nigra compacts (SNc) and striatum in the normal side of the rat model of PD were decreased (P < 0.05), which proved the successful establishment of PD models. The NAA/Cr ratio of the injected side striatum of model group was lower than that of the normal side (P < 0.05). The ratios of Cho/Cr showed no significant difference between the two sides (P > 0.05). These results indicated that the modified 1.5T 1H-MRS should be a noninvasive technique which could provide useful information about the biochemical metabolites in striatum for the study of PD in rat model.
Animals ; Corpus Striatum ; enzymology ; Female ; Magnetic Resonance Spectroscopy ; methods ; Male ; Oxidopamine ; Parkinson Disease, Secondary ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tyrosine 3-Monooxygenase ; metabolism

The somatic cells can be induced into ES-like stem cells when retrovirally infected the defined transcription factors including Oct4, Sox2, Klf4 and c-Myc. These ES-like cells are named induced pluripotent stem (iPS) cells and this method is called iPS technology. Until the end of 2009, iPS cell lines have been generated in various animal species, such as mouse, human, rhesus monkey, rat and pig. Mouse iPS cells are also used to generate chimera mice and viable mice through the tetraploid complementation. Although iPS cells are extremely similar to ES cells in both morphology and growth features, to generate iPS cells do need the defined culture procedures. Based on the update global iPS technology development and the iPS studies in our laboratory, this paper focused on the establishment of iPS cell lines and improvement of iPS cell culture condition.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; Embryonic Stem Cells ; cytology ; metabolism ; Humans ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; SOXB1 Transcription Factors ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

Phospholipase D (PLD) hydrolyzes phosphocholine into choline and phosphatide acid, and these metabolites play an important role in regulating cell physiology and biochemistry. To study the biological function of phospholipase D3 (PLD3) during the insulin stimulation in C2C12 myoblasts, we constructed PLD3 over-expressed cell lines (C2C12/pPLD3) and investigated the phosphorylation of Akt. The results showed that the level of phosphorylated Akt (P-Akt) was significantly increased in control C2C12 cells when insulin concentration was elevated during cell treatment, whereas the level of P-Akt in C2C12/pPLD3 cells was not changed. When extending the time of insulin treatment, P-Akt level in C2C12/pPLD3 cells was increased around 2 folds, but the total level of P-Akt in C2C12/pPLD3 was still lower than that in control group. 1-Butanol, a PLD inhibitor, could completely block Akt phosphorylation in C2C12 cells that even stimulated by insulin. However, 1-Butanol did not inhibit the Akt phosphorylation in C2C12/pPLD3 cells, but increased the phosphorylation up to 6 folds higher than control cells. The level of Akt phosphorylation in control C2C12 cells was increased significantly when stimulated by phosphatidic acid (PA), while there was no change in C2C12/pPLD3 cells with the similar treatment. When cells simulated by both PA and insulin, P-Akt level in both C2C12/pPLD3 cells and C2C12 cells were down regulated. Our observations indicated that PLD3 over expression may inhibit Akt phosphorylation and further block the transduction of insulin signaling in C2C12 cells.
Cell Line ; Humans ; Insulin ; pharmacology ; Myoblasts ; cytology ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phospholipase D ; biosynthesis ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; chemistry ; drug effects ; Signal Transduction

Pregnancy and childbirth are natural physiological processes for women, but most mothers, especially primipara, tend to suffer from anxiety, fear and other unhealthy psychology, accompanied by the occurrence of these bad emotions. It is a worthy concern that whether the subjective well-being of pregnant women will be affected by these bad emotions. However, there are few reports about subjective well-being of pregnant women in China. In this study, the subjective well-being′s concept, assessment tools and influencing factors of maternal subjective well-being were reviewed. The paper also described the research progress and development on the subjective well-being of pregnant women. The purpose of this paper is to provide reference for the study and practice of in China.