Objective: To investigate the function and mechanism of phospholipase Cγ1 (PLCγ1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-κB) were used to study the effect of PLCγ1 and NF-κB on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLCγ1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLCγ1 or NF-κB resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P<0.05), but had no marked effect on SW480 cells. Western blot analysis showed that epidermal growth factor (EGF) stimulated the phosphorylation of PLCγ1 in LoVo. The results of EMSA indicated that inhibition of PLCγ1 signaling pathway also down-regulated the activity of NF-κB while EGF reversed the function. Conclusion:These data suggest that PLCγ1 plays a pivotal role in the EGF-induced cell-matrix adhesion of highly metastatic colorectal cancer cells and that NF-κB is also functional in this signaling pathway.

Objective: To investigate the function and mechanism of phospholipase C?1 (PLC?1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-?B) were used to study the effect of PLC?1 and NF-?B on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLC?1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLC?1 or NF-?B resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P

On the base of element and metabolism balancing,the mathematic model of the human-like collagen expression phase with recombinant Escherichia coli BL21 was developed and the unknown parameters in the model were estimated with the method of nonlinear optimization.The model was in agreement with the growth kinetics and the metabolic kinetics,and the key calculated parameters of ?h,?p and mx were 1.173 mol?C-mol-1,293.814 mol?C-mol-1 and 17.878 mol?C-mol-1?h-1 respectively.This model could preferably predict the macroscopic reaction rates,and in the synthesis phase of human-like collagen,the specific growth rate should be controlled at 0.04 h-1 with controlling glucose feeding rate to gain the highest specific production rate of human-like collagen.

To investigate dynamic characteristics of the hydrolization lipid phosphatidylinositol(4,5)-bisphosphate(PIP 2 )with the stimulation of epidermal growth factor(EGF),the mathematical model to simulate the metabolizability of PIP 2 is established based on Law of Mass Action and Law of the Quasi-steady-State Approximation.Differential equations of concentration relations between PIP 2 and EGF receptor are formulated,and the effect of the parameters on the changing trend of PIP 2 is analyzed.This mathematical model describes biology characteristics of metabolized PIP 2 and dependency relationships of concentration between key signal producuts.

BACKGROUND:Spinocerebel ar ataxia type 3 (SCA3) is a typical genetic neurodegenerative disease. To establish patient-specific disease models of genetic background contributes to studying the pathogenesis and exploring therapeutic manners.
OBJECTIVE:To observe the effectiveness of neural differentiation of induced pluripotent stem cel lines induced by SCA3 and the stability of CAG copy number.
METHODS:Skin tissue of SCA3 patient was obtained clinical y, and specific skin flbroblasts were isolated and cultured. Reprogramming fibroblasts could obtain induced pluripotent stem cel s. Patient-specific induced pluripotent stem cel s, normal person induced pluripotent stem cel s (NHF) and embryonic stem cel s (ES-10) were induced to differentiate. Flow cytometry was used to compare the efficiency of differentiation. Western blot assay was utilized to detect ataxin-3 protein expression in neurons. Polymerase chain reaction was applied to measure the CAG repeat number of SCA3/ATXN3 gene.
RESULTS AND CONCLUSION:Induced pluripotent stem cel s that had identical genetic background to fibroblasts were successful y obtained, and had similar morphology and multi-directional differentiation potential to human embryonic stem cel s. Each cel line could differentiate into neural stem cel s. The CAG number did not apparently alter before and after reprogramming as wel as induction of neuronal differentiation. The effectiveness of the differentiation of induced pluripotent stem cel s derived from SCA3 into neural stem cel s was lower than that of normal person-derived induced pluripotent stem cel s (NHF) and embryonic stem cel s (ES-10). These findings demonstrate that reprogramming can successful y establish human induced pluripotent stem cel s, and induced the differentiation of above cel s into neural stem cel s. In the whole process, CAG number did not obviously alter, which was consistent with body cel s of patients.

Objective: To investigate the function and mechanism of phospholipase Cγ1 (PLCγ1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-κB) were used to study the effect of PLCγ1 and NF-κB on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLCγ1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLCγ1 or NF-κB resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P<0.05), but had no marked effect on SW480 cells. Western blot analysis showed that epidermal growth factor (EGF) stimulated the phosphorylation of PLCγ1 in LoVo. The results of EMSA indicated that inhibition of PLCγ1 signaling pathway also down-regulated the activity of NF-κB while EGF reversed the function. Conclusion:These data suggest that PLCγ1 plays a pivotal role in the EGF-induced cell-matrix adhesion of highly metastatic colorectal cancer cells and that NF-κB is also functional in this signaling pathway.

Animals ; Cardiomegaly ; Constriction ; Heart ; Mice

Objective To prepare a novel dual-modality imaging probe based on Cerasome nano-materials, and evaluate its in vivo biodistribution and pharmacokinetic properties. Methods ICG encapsu-lated Cerasome was modified with chelating agent DOTA for 111 In-labeling. Normal mice firstly were used for in vivo studies. Animals were sacrificed at different time points after tail vein administration, blood samples were taken and the organs of interest were captured to evaluate the pharmacokinetic properties and in vivo biodistribution of 111 In-ICG-DPDCs. The subcutaneous Lewis lung carcinoma ( LLC ) tumor model in C57BL/6 mouse was established. The tumor-bearing mice were subjected to optical imaging in small animal IVIS and SPECT imaging in small animal nanoScanSPECT/CT system for tumor uptake of 111 In-ICG-DPDCs. Results The size of the nanoparticle probe was about 90 nm, and the 111 In-labeling was successfully per-formed with 99.93% radiochemical purity after purification. 111 In-ICG-DPDCs showed excellent in vitro sta-bility with 97.10% radiochemical purity at 48 h post-purification. In vivo blood clearance experiments showed that 111 In-ICG-DPDCs had a relative long blood circulation time with the fast and slow phase half-lives of 40 and 132.7 min. 111In-ICG-DPDCs accumulated mainly in the liver and spleen, with long retention time. NanoScanSPECT/CT imaging showed that LLC tumors were significantly visualized at 4 h post-injection, and the other major accumulated organs were the liver and spleen, which were consistent with the results of biodistribution. Optical imaging showed significant uptake of the nanoparticle probe in the tumor, confirming the SPECT imaging results. Conclusion The Cerasome based probe designed could be used for tumor SPECT and optical dual-modality imaging, and has potential for therapeutic use.

OBJECTIVETo express human platelet-derived growth factor (hPDGF) B chain mature peptide gene in a prokaryotic expression system and detect the bioactivity of the expressed protein.

METHODShPDGF B chain mature peptide gene was amplified and expressed in E. coli, and the recombinant protein, rhPDGF-BB, was purified and renatured in GSSG/GSS system. The bioactivity of rhPDGF-BB in vitro was evaluated with SD rat osteoblasts.

RESULTSThe full-length PDGF-B mature peptide gene was obtained and verified, and successfully expressed in E. coli. Bioactivity detection results showed that the expressed rhPDGF-BB obviously promoted the proliferation and DNA replication of SD rat osteoblasts in vitro (P<0.01).

CONCLUSIONhe PDGF-B chain mature peptide cDNA has been successfully cloned and the PDGF-B precursor highly expressed in E. coli, and renatured rhPDGF-BB displays high bioactivity as shown by MTT assay and flow cytometry. This success provides the basis for production of functional PDGF-BB and facilitates further studies of its role in fracture healing and trauma reconstruction.

Animals ; Cell Proliferation ; Cells, Cultured ; DNA Replication ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Osteoblasts ; metabolism ; Proto-Oncogene Proteins c-sis ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; genetics

Objective: To investigate the efficacy and safety of Rivaroxaban for elderly patients with thrombotic diseases. Methods: This was a retrospective study.A total of 301 elderly patients taking Rivaroxaban from October 2012 to November 2017 at the Second Medical Center of the Chinese PLA General Hospital were consecutively selected.The ages ranged from 60 to 102 years, with an average age of(86.5±8.4)years.Anticoagulation regimens were developed based on comprehensive evaluation of indications, creatinine clearance, ischemia and bleeding risk.Patients were divided into a Rivaroxaban 2.5-5.0 mg/d group(n=72), a 10.0 mg/d group(n=205), and a 15.0-20.0 mg/d group(n=24). Hepatic function, renal function, and coagulation indexes were measured before and after the administration of Rivaroxaban.Fatal bleeding, cardiovascular deaths, all-cause deaths, non-fatal bleeding and thromboembolic events were recorded during the follow-up period. Results: The average dose of Rivaroxaban was(9.3±3.0)mg/d, and the minimum dose was 2.5 mg/d.The average follow-up time was(14.9± 13.9)months and the longest follow-up time was 48 months.One patient had intracranial bleeding.Twenty patients(6.6%)died with a cumulative incidence of 25.2%, three(1.0%)died of cardiac events, and 55.0% died of pneumonia and multiple organ failure.Forty patients(13.3%)had non-fatal hemorrhagic events with a cumulative incidence of 42.4%.Seven patients(2.3%)had thromboembolic events with a cumulative incidence of 16.0%, including 2 cases of non-fatal myocardial infarction, 3 cases of cerebral infarction and 2 cases of deep vein thrombosis.After treatment, levels of prothrombin time and fibrinogen significantly increased while levels of D-dimer significantly deceased(P<0.05). Conclusions Compared with previous reports, low-dose Rivaroxaban is safe and effective for elderly patients with thrombotic diseases.However, the risk of bleeding and ischemia should be comprehensively evaluated, and appropriate doses of Rivaroxaban should be selected individually.