ObjectiveTo summarize the latest progress of parathyroid gland identification in thyroid surgery, and to provide some reference for improving the clinical efficacy.MethodThe literatures about the identification of parathyroid gland in thyroid surgery in recent years were collected to make an review.ResultsThere were many methods for identifying parathyroid gland in thyroid surgery, such as naked eye identification method, intraoperative frozen section, intraoperative staining identification method, intraoperative optical identification method, intraoperative parathyroid hormone assay, γ-detector, and histological identification, each method had its own advantages and disadvantages.ConclusionThe identification of parathyroid gland does not only depend on a certain method, but also require surgeons to enhance their ability to distinguish parathyroid gland.
Acute respiratory distress syndrome (ARDS) is a serious threat to human life and health disease, with acute onset and high mortality. The current diagnosis of the disease depends on blood gas analysis results, while calculating the oxygenation index. However, blood gas analysis is an invasive operation, and can’t continuously monitor the development of the disease. In response to the above problems, in this study, we proposed a new algorithm for identifying the severity of ARDS disease. Based on a variety of non-invasive physiological parameters of patients, combined with feature selection techniques, this paper sorts the importance of various physiological parameters. The cross-validation technique was used to evaluate the identification performance. The classification results of four supervised learning algorithms using neural network, logistic regression, AdaBoost and Bagging were compared under different feature subsets. The optimal feature subset and classification algorithm are comprehensively selected by the sensitivity, specificity, accuracy and area under curve (AUC) of different algorithms under different feature subsets. We use four supervised learning algorithms to distinguish the severity of ARDS (P/F ≤ 300). The performance of the algorithm is evaluated according to AUC. When AdaBoost uses 20 features, AUC = 0.832 1, the accuracy is 74.82%, and the optimal AUC is obtained. The performance of the algorithm is evaluated according to the number of features. When using 2 features, Bagging has AUC = 0.819 4 and the accuracy is 73.01%. Compared with traditional methods, this method has the advantage of continuously monitoring the development of patients with ARDS and providing medical staff with auxiliary diagnosis suggestions.
Objective
To establish a simple and efficient method to isolate and culture the umbilical vein vascular endothelial cells in canine.
Methods
Twelve umbilical cords [(13.0 ± 1.5) cm in length] were taken from 12 newborn pups of Beagles. And then the vascular endothelial cells were isolated from these umbilical cords digested by 1% collagenase type I for 5, 7, and 10 minutes respectively (4 umbilical cords in each group). After cultured, the vascular endothelial cells were identified by morphology, immunofluorescence, and flow cytometry. And the growth curvature of umbilical vein vascular endothelial cells was detected by MTT assay.
Results
Few vascular endothelial cells were collected at 5 and 10 minutes after digestion; many vascular endothelial cells were seen at 7 minutes, and became cobblestone with culture time, with a large nucleus; after passage, cell morphology had no obvious change. Fluorescence microscope results showed that positive von Willebrand factor (vWF) and CD31 cells were observed in most of cells. The flow cytometry test displayed that the positive cell rates of vWF and CD31 were 99.0% ± 0.7% and 98.0% ± 1.2%, respectively. The above results indicated that cultured cells were vascular endothelial cells. MTT assay showed that vascular endothelial cells proliferation increased significantly with culture time.
Conclusion
Enzyme digestion is a convenient method to isolate vascular endothelial cells from canine umbilical vein, and a large number of cells and high purity of cells can be obtained by the method.
Multi-layer perceptron (MLP) neural network belongs to multi-layer feedforward neural network, and has the ability and characteristics of high intelligence. It can realize the complex nonlinear mapping by its own learning through the network. Bipolar disorder is a serious mental illness with high recurrence rate, high self-harm rate and high suicide rate. Most of the onset of the bipolar disorder starts with depressive episode, which can be easily misdiagnosed as unipolar depression and lead to a delayed treatment so as to influence the prognosis. The early identification of bipolar disorder is of great importance for patients with bipolar disorder. Due to the fact that the process of early identification of bipolar disorder is nonlinear, we in this paper discuss the MLP neural network application in early identification of bipolar disorder. This study covered 250 cases, including 143 cases with recurrent depression and 107 cases with bipolar disorder, and clinical features were statistically analyzed between the two groups. A total of 42 variables with significant differences were screened as the input variables of the neural network. Part of the samples were randomly selected as the learning sample, and the other as the test sample. By choosing different neural network structures, all results of the identification of bipolar disorder were relatively good, which showed that MLP neural network could be used in the early identification of bipolar disorder.
ObjectiveTo compare the biological features of early and late endothelial progenitor cells (EPCs) by isolating and culturing early and late EPCs from the human peripheral blood so as to find some unique properties of EPCs and to propose a suitable strategy for EPCs identification.
MethodsMononuclear cells were isolated from the human peripheral blood using density gradient centrifugation. Then, the cells were inoculated in human fibronectin-coated culture flasks and cultured in endothelial cell basal medium 2. After 4-7 days and 2-3 weeks culture, early and late EPCs were obtained respectively. The morphology, proliferation potential, surface markers, cytokine secretion, angiogenic ability, and nitric oxide (NO) release were compared between 2 types of EPCs. Meanwhile, the human aortic endothelial cells (HAECs) were used as positive control.
ResultsThe morphology of early and late EPCs was different:early EPCs formed a cell cluster with a spindle shape after 4-7 days of culture, and late EPCs showed a cobblestone appearance. Late EPCs were characterized by high proliferation potential and were able to form capillary tubes on Matrigel, but early EPCs did not have this feature. Both types EPCs could ingest acetylated low density lipoprotein and combine with ulex europaeus Ⅰ. Flow cytometry analysis showed that early EPCs did not express CD34 and CD133, but expressed the CD14 and CD45 of the hematopoietic stem cell markers;however, late EPCs expressed CD31 and CD34 of the endothelial cell markers, but did not express CD14, CD45, and CD133. By RT-PCR analysis, the expressions of vascular endothelial growth receptor 2 and vascular endothelial cadherin in early EPCs were significantly lower than those in the late EPCs and HAECs (P<0.05), but no significant difference was found in the expression of von Willebrand factor and endothelial nitric oxide synthase (eNOS) between 2 type EPCs (P>0.05). The concentrations of vascular endothelial growth factor, granulocyte colony-stimulating factor, and interleukin 8 were significantly higher in the supernatant of early EPCs than late EPCs (P<0.05). Western blot assay indicated eNOS expressed in both types EPCs, while the expression of eNOS in late EPCs was significantly higher than early EPCs at 5 weeks (P<0.05). Both cell types could produce similar amount of NO (P>0.05).
ConclusionThe expression of eNOS and the production of NO could be used as common biological features to identify EPCs, and the strategy of a combination of multiple methods for EPCs identification is more feasible.
OBJECTIVE: To investigate the selection and identification of human keratinocyte stem cells(KSC) in vitro. METHODS: According to the characteristics of KSC which can adhere to extracellular matrix very fast, we selected 3 groups of different time(5 minutes, 20 minutes and 60 minutes) and unselected as control group. And the cells were identified by monoclone antibody of beta 1-integrin and cytokeratin 19 (Ck19), then the image analysis was done. Furthermore we analyzed the cultured cells with flow cytometer(FCM) and observed the ultrastructure of the cell by transmission electron microscope(TEM). RESULTS: The cell clones formed in all groups after 10 to 14 days, while the cells of 5 minute group grew more slowly than those of the other groups, however, the clones of this group were bigger. The expression of beta 1-integrin and Ck19 were found in all groups. The positive rate of beta 1-integrin was significant difference between 5 minute group and the other groups (P lt; 0.05). And the expression of Ck19 was no significant difference between 5 minute group and 20 minute group(P gt; 0.05), and between 60 minute group and control group. But significant difference was observed between the former and the later groups(P lt; 0.05). The result of FCM showed that most cells of the 5 minute group lied in G1 period of cell cycle, which was different from those of the other groups. At the same time, the cells of 5 minute group were smaller and contained fewer organelles than those of the other groups. CONCLUSION: The above results demonstrate that the cells of 5 minute group have a slow cell cycle, characteristics of immaturity, and behaving like clonogenic cells in vitro. The cells have the general anticipated properties for KSC. So the KSC can be selected by rapid attachment to extracellular matrix and identified by monoclone antibody of beta 1-integrin and Ck19.
Intravascular ultrasound (IVUS) is widely used in coronary artery examination. Ultrasonic elastography combined with IVUS is very conspicuous in identifying plaque component and in detecting plaque vulnerability degree. In this study, a simulation model of the blood vessel based on finite element analysis (FEA) was established. The vessel walls generally have radial changes caused by different intravascular pressure. The signals at lower pressures were used as the pre-deformation data and the signals at higher pressure were used as the post-deformation data. Displacement distribution was constructed using the time-domain cross-correlation method, and then strain images. By comparison of elastograms under different pressures, we obtained the optimal pressure step. Furthermore, on the basis of the obtained optimize pressure step, the simulation results showed that this method could effectively distinguish characteristics between different component plaques, and could guide the later experiments and clinical applications.
Esophageal carcinoma is a kind of common malignant tumor in the digestive tract. Although a lot of basic researches and clinical trials have been carried out all over the world, neither the diagnostic level nor the therapeutic effects have been obviously improved. The 5-year survival rate of esophageal cancer patient is still lower than that of other malignant tumors. Up to now, some frontier researches consider that the reason of the esophageal carcinoma being difficult to cure is related to the stem cells in it. Elimination or suppression of these stem cells may bring new hope for the treatment of esophageal cancer. This article generally introduces the specific markers, separation and indentification methods about the esophageal cancer stem cell. The targeted therapy is also mentioned.
Objective
To establish a method for quality control of Astragalus Radix and Scutellariae Radix in Biqiaotong granules and provide basis for the establishment of quality standard.
Methods
The single-factor test method was used to investigate the factors of thin layer chromatography (TLC) conditions, including different extract method and solvents, developing system, comogemc agents, temperature, humidity, drawing amounts and thin layer boards, and to screen the best TLC conditions of Astragalus Radix and Scutellariae Radix .
Results
The TLC conditions of Astragalus Radix were used trichloromethane-methanel-water (13:7:2) as developing solvent, separated on silica gel G, heatd under 105℃ until the spots bacame clear. The TLC conditions of Scutellariae Radix were used methylbenzene-ethy acetate- formic acid-methanel (9:3:2:2) as developing solvent, separated on silica gel G, observed after 30 minutes under daylight until the spots were clear.
Conclusions
The spot features are clear, and with good separating degree, strong specificity, and good repeatability without the inference of negative control. The TLC method is simple, sensitive and accurate, which can be adopted for the quality control of Biqiaotong granules.
ObjectiveTo explore the methods of separation， culture， and identification of breast cancer stromal fibroblasts (BCSFs), which could build up a good basis for the further research of function. MethodsBreast cancer tissues were obtained during breast cancer operation， and were cut into pieces with size of 1 mm×1 mm×1 mm under aseptic conditions， then the pieces of the tissues were digested by collagenase Ⅰ and hyaluronidase. Finally the cells separated from the tissues incubated at 37 ℃ with 5% CO2 and 95% air humidified incubator. Morphological characteristics of the fibroblasts were observed under light microscope. The certain proteins were examined by immunohistochemistry （using CK， Vimentin， α-SMA, and TE-7 antibody） and flow cytometric analysis (CD34 and CD45). ResultsThe separated cells begin to attach to the wall of flask within 24 h and reached almost confluency in about 7 d to 10 d . According to identification, the successful rate of separation and culture of BCSFs was 90%（18/20）, and the characteristics of cells showed that morphological characteristics of the fibroblasts was flat spindle， rich cytoplasm, and a flat ovoid cystic nuclear. The fibroblasts in breast cancer tissues showed negative staining for cytokeratin， positive staining for vimentin， alpha-smooth muscle actin, and TE-7， and negative for CD34 and CD45 by flow cytometric analysis. ConclusionsThe fibroblasts in breast cancer tissues could be easily obtained by tissues cuting combined enzyme digestion and rocking technology in vitro. The present study provide an experimental foundation for further studies on fibroblasts in breast cancer.
