ObjectiveTo detect the expressions of takeda G protein-coupled receptor 5 (TGR5) and mortalin protein 75 in the tissues of patients with intrahepatic cholangiocarcinoma (ICC), and to explore their relationship with prognosis.MethodsA total of 94 ICC patients who were admitted to Anyang District Hospital and received surgical treatment from March 2015 to March 2018 were selected as the research objects. The expressions of TGR5 and mortalin protein 75 in ICC cancer tissues and adjacent tissues were detected by immunohistochemistry and Western blot (WB). The relationship between the expressions of TGR5 and mortalin protein 75 in ICC cancer tissues and clinicopathological parameters and prognosis was analyzed. Multivariate Cox proportional hazards regression was used to analyze the risk factors of poor prognosis in patients with ICC. ROC curve was used to analyze the diagnostic value of TGR5 and mortalin protein 75 for poor prognosis in patients with ICC.ResultsImmunohistochemical results showed that the positive expression rates of TGR5 and mortalin protein 75 in cancer tissues were significantly higher than those in adjacenttissues (P<0.05). WB results showed that the protein expression levels of TGR5 and mortalin protein 75 in cancer tissues were significantly higher than those in adjacent tissues (P<0.05). The expression of TGR5 protein in cancer tissues of ICC patients was correlated with gender, tumor diameter, degree of differentiation, TNM staging, satellite focus, and liver cirrhosis (P<0.05). The expression of mortalin protein 75 was correlated with tumor diameter, TNM staging, nerve involvement, satellite focus, and liver cirrhosis (P<0.05). There were significant differences in gender, tumor diameter, TNM staging, microvascular invasion, satellite focus, liver cirrhosis, and the expressions of TGR5 and mortalin protein 75 between the poor prognosis group and the good prognosis group (P<0.05). The cumulative 3-year overall survival rate of TGR5 positive patients (32.00%) was significantly lower than that of TGR5 negative patients (63.16%), χ2=6.228, P=0.013; the cumulative 3-year overall survival rate of mortalin protein 75 positive patients (32.91%) was significantly lower than that of mortalin protein 75 negative patients (66.67%), χ2=6.079, P=0.014. Multivariate Cox proportional hazards regression analysis showed that the positive expression of TGR5 and mortalin protein 75, TNM Ⅲ+Ⅳphase, satellite focus, and cirrhosis were risk factors for poor prognosis in ICC patients (P<0.05). ROC results showed that when the expression level of TGR5 was 0.932 as the cut-off value, its AUC in the diagnosis of poor prognosis of ICC patients was 0.783, the sensitivity was 72.4%, the specificity was 72.2%; when the expression level of mortalin protein 75 was 0.756 as the cut-off value, its AUC in the diagnosis of poor prognosis of ICC patients was 0.805, the sensitivity was 84.4%, the specificity was 63.9%; the AUC of combined diagnosis of TGR5 and mortalin protein 75 was 0.884, the sensitivity was 79.3%, the specificity was 83.3%.ConclusionsThe high expressions of TGR5 and mortalin protein 75 in cancer tissues of ICC patients are associated with poor prognosis, and they are risk factors for poor prognosis. The combined detection of TGR5 and mortalin protein 75 has a certain value in predicting poor prognosis, and can be used as potential biological indicators.
Ras homolog family (Rho)/ Rho-associated coiled-coil kinase (ROCK) signaling pathway widely exists in human and mammal cells, which is closely related to inhibition of repair after optic nerve damage. The expression level of Rho/ROCK signaling pathway-related proteins is up-regulated in glaucoma, and related with the death of retinal ganglionic cell (RGC) and the axon activity. ROCK inhibitors can protect the surviving RGC and promote axon extension with a dose-dependent manner. ROCK inhibitors also can inhibit glial scar formation, lower intraocular pressure and inhibit inflammatory response to some degrees. Rho/ROCK signaling pathway correlates with the optic nerve disease progression, and ROCK inhibitors hope to become a new therapeutic drug.
In order to detect the flow characteristics of blood samples in the capillary, this paper introduces a blood flow velocity measurement system based on field-programmable gate array (FPGA), linear charge-coupled devices (CCD) and personal computer (PC) software structure. Based on the analysis of the TCD1703C and AD9826 device data sheets, Verilog HDL hardware description language was used to design and simulate the driver. Image signal acquisition and the extraction of the real-time edge information of the blood sample were carried out synchronously in the FPGA. Then a series of discrete displacement were performed in a differential operation to scan each of the blood samples displacement, so that the sample flow rate could be obtained. Finally, the feasibility of the blood flow velocity detection system was verified by simulation and debugging. After drawing the flow velocity curve and analyzing the velocity characteristics, the significance of measuring blood flow velocity is analyzed. The results show that the measurement of the system is less time-consuming and less complex than other flow rate monitoring schemes.
In order to overcome the difficulty in lung parenchymal segmentation due to the factors such as lung disease and bronchial interference, a segmentation algorithm for three-dimensional lung parenchymal is presented based on the integration of surfacelet transform and pulse coupled neural network (PCNN). First, the three-dimensional computed tomography of lungs is decomposed into surfacelet transform domain to obtain multi-scale and multi-directional sub-band information. The edge features are then enhanced by filtering sub-band coefficients using local modified Laplacian operator. Second, surfacelet inverse transform is implemented and the reconstructed image is fed back to the input of PCNN. Finally, iteration process of the PCNN is carried out to obtain final segmentation result. The proposed algorithm is validated on the samples of public dataset. The experimental results demonstrate that the proposed algorithm has superior performance over that of the three-dimensional surfacelet transform edge detection algorithm, the three-dimensional region growing algorithm, and the three-dimensional U-NET algorithm. It can effectively suppress the interference coming from lung lesions and bronchial, and obtain a complete structure of lung parenchyma.
ObjectiveTo investigate the mechanism of G protein coupled receptor kinase interacting protein 1 (GIT1) affecting angiogenesis by comparing the differentiation of bone marrow mesenchymal stem cells (BMSCs) differentiated into endothelial cells between GIT1 wild type mice and GIT1 gene knockout mice.MethodsMale and female GIT1 heterozygous mice were paired breeding, and the genotypic identification of newborn mice were detected by PCR. The 2nd generation BMSCs isolated from GIT1 wild type mice or GIT1 gene knockout mice were divided into 4 groups, including wild type control group (group A), wild type experimental group (group A1), GIT1 knockout control group (group B), and GIT1 knockout experimental group (group B1). The cells of groups A1 and B1 were cultured with the endothelial induction medium and the cells of groups A and B with normal cluture medium. The expressions of vascular endothelial growth factor receptor 2 (VEGFR-2), VEGFR-3, and phospho-VEGFR-2 (pVEGFR-2), and pVEGFR-3 proteins were detected by Western blot. The endothelial cell markers [von Willebrand factor (vWF), platelet-endothelial cell adhesion molecule 1 (PECAM-1), and vascular endothelial cadherin (VE-Cadherin)] were detected by flow cytometry. The 2nd generation BMSCs of GIT1 wild type mice were divided into 4 groups according to the different culture media: group Ⅰ, primary cell culture medium; group Ⅱ, cell culture medium containing SAR131675 (VEGFR-3 blocker); group Ⅲ, endothelial induction medium; group Ⅳ, endothelial induction medium containing SAR131675. The endothelial cell markers (vWF, PECAM-1, and VE-Cadherin) in 4 groups were also detected by flow cytometry.ResultsWestern blot results showed that there was no obviously difference in protein expressions of VEGFR-2 and pVEGFR-2 between groups; and the expressions of VEGFR-3 and pVEGFR-3 proteins in group A1 were obviously higher than those in groups A, B, and B1. The flow cytometry results showed that the expressions of vWF, PECAM-1, and VE-Cadherin were significantly higher in group A1 than in groups A, B, and B1 (P<0.05), and in group B1 than in groups A and B (P<0.05); but no significant difference was found between groups A and B (P>0.05). In the VEGFR-3 blocked experiment, the flow cytometry results showed that the expressions of vWF, PECAM-1, and VE-Cadherin were significantly higher in group Ⅲ than in groupsⅠ, Ⅱ, and Ⅳ, and in group Ⅳ than in groups Ⅰ and Ⅱ (P<0.05); but no significant difference was found between groups Ⅰ and Ⅱ (P>0.05).ConclusionGIT1 mediates BMSCs of mice differentiation into endothelial cells via VEGFR-3, thereby affecting the angiogenesis.
Aiming at long signal acquisition time, low flux, bad signal-to-noise ratio and low intelligence in coloration biochip reader, a new kind of rapid device with high flux was developed. The device consisted of hardware system and software system. It used a charge-coupled device (CCD) as the photoelectric sensor elements and obtained the biochip microarray image. The device integrated the embedded operating system based on i.MX6 chip. The microarray image processing, data analysis and result output were achieved through the code information of the software chip. Experiments with the standard grayscale sheet and standard format chip were carried out. The results showed that the maximum measurement error was less than 0.1%, the value of R2 was 98.7%, and the value of CV was 1.096 1%. The comparison results of 200 samples showed that detection performance of the proposed device was better than that of the same kind of marketed equipment.
Breast cancer patients and their spouses face a series of problems in their sexual life, which together lead to the decline of the sexual life quality of the couple, endangering the physical and mental health and the marital relationship of the patients and their spouses. Therefore, this article reviews the current situation, assessment tools, and influencing factors of sexual life quality of breast cancer patients and their spouses, and introduces the foreign intervention measures of sexual life quality based on the dual perspective of husband and wife, in order to promote the development of research on sexual life of breast cancer patients and their spouses in China, and provide references for the construction of a localized intervention program to improve the sexual life quality.
Objective To study the function and mechanism of G protein coupled receptor kinase interacting protein 1(GIT1) RNA hairpin (GIT1-RNAh) in osteoblast migration. Methods The sixth passage osteoblasts were divided into 2 groups and were infected by GIT1-RNAh (experimental group) and green fluoresence protein RNA hairpin (GFP-RNAh) (control group) adenovirus for 12 hours respectively. Each group was further classfied into two groups according to with or without platelet-drived growth factor (PDGF) stimulation. The GIT1 expression and Paxillindistribution was analyzed by immunofluorescence staining. Paxillin phosphorylation was detected by Western Blot. The localization of Paxillin was determined by co-immunofluorescence staining after transfection with cyanine fluorescence protein tagged GIT1RNAh (CFP-GIT1-RNAh)(experimental group) and GFP-RNAh (CFP-GFP-RNAh)(control group). The role of GIT1-RNAh (experimental group) and GFP-RNAh (control group) adenovirus in osteoblasts migration was determined by wound healing assay. Results Immunofluorescence staining results showed that the GIT1-RNAh significantly inhibited endogenous GIT1 expression, interfered Paxillin distribution.Western Blot results showed that Paxillin phosporylation was obviously inhibited in osteoblasts infected with GIT1-RNAh adenovirus (P＜0.05). The wound healing assay results howed that GIT1-RNAh adenovirus significantly inhibited osteoblast migration induced by PDGF. Conclusion GIT1-RNAh inhibits osteoblasts migration by interfering paxillin distribution and decrease Paxillin phosphorylation.
ObjectiveTo investigate the effects and mechanisms of G protein-coupled receptor 91 (GPR91) on blood-retinal barrier (BRB) in diabetic rats. MethodsA lentiviral vector of shRNA targeting rat GPR91 and scrambled shRNA were constructed. Healthy male Sprague-Dawley (SD) rats were selected in this study. The 60 rats were randomized into 4 groups and treated as follows:(1) control group (Group A, n=15), the rats received injections of an equal volume of 0.1% citrate buffer; (2) streptozocin (STZ) group (Group B, n=15), the rats received injections of STZ; (3) LV.shScrambled group (Group C, n=15), diabetic rats received an intravitreal injection of 1 μl 1×108 TU/ml scrambled shRNA lentiviral particles at 2 weeks after the induction of diabetes; (4) LV.shGPR91 group (Group D, n=15), diabetic rats received an intravitreal injection of 1 μl 1×108 TU/ml pGCSIL-GFP-shGPR91 lentiviral particles. At 12 weeks after intravitreal injection, immunohistochemistry and Western blot were used to assess the expression of GPR91, p-extracellular signal-regulated kinase(ERK)1/2, t-ERK1/2, p-Jun N-terminal kinase (JNK), t-JNK, p-p38 mitogen-activated protein kinase (MAPK) and t-p38 MAPK. Haematoxylin and eosin (HE) staining and Evans blue dye were used to assess the structure and function of the retinal vessel. Immunohistochemistry enzyme-linked immunosorbent assay (ELISA) was used to test the protein level of VEGF. ResultsImmunohistochemistry staining showed that GPR91 was predominantly localized to the cell bodies of the ganglion cell layer. Western blot showed that GPR91 expression in Group D decreased significantly compared with Group C (F=39.31, P < 0.01). HE staining showed that the retina tissue in Group B and C developed telangiectatic vessels in the inner layer of retina, while the telangiectatic vessels attenuated in Group D. It was also demonstrated in Evans blue dye that the microvascular leakage in Group D decreased by (33.8±4.11)% compared with Group C and there was significant difference (F=30.35, P < 0.05). The results of ELISA showed the VEGF secretion of Group B and C increased compared with Group A and the VEGF expression in Group D was significantly down regulated after silencing GPR91 gene (F=253.15, P < 0.05).The results of Western blot indicated that compared with Group A, the expressions of p-ERK1/2, p-JNK and p-p38 MAPK were significantly upregulated (q=6.38, 2.94, 3.45;P < 0.05). Meanwhile, the activation of ERK1/2 was inhibited by GPR91 shRNA and the difference was statistically significant (F=22.50, P < 0.05). ConclusionsThe intravitreal injection of GPR91 shRNA attenuated the leakage of BRB in diabetic rats. GPR91 regulated the VEGF release and the leakage of BRB possibly through the ERK1/2 signaling pathway.
The mechanisms of general anesthesia, which was introduced about 170 years ago, remain poorly understood. Even less well understood are the effects of general anesthesia on the human body. Recently we identified 18 G-protein coupled receptor (GPCR) genes of Daphnia pulex, an invertebrate model organism. Phylogenetic analysis identified these genes to be the homologs of the human γ-aminobutyric acid, type B (GABAB) receptor, metabotropic glutamate receptors (mGluR), adrenergic receptor, serotonin (5-HT) receptor, dopamine receptor and muscarinic acetylcholine receptor (mAChR). Using reverse transcription and quantitative PCR techniques, we systematically measured the effects of propofol, etomidate and ethanol on these 18 GPCR mRNA expressions in Daphnia pulex.