ABSTRACT
Objective To establish a rat model of post-traumatic stress disorder(PTSD)through single-prolonged stress(SPS)and to observe the effect of social isolation on the behavior of the SPS model rats. Methods A total of 36 rats were randomly divided into the normal group, SPS model group and SPS combined with social isolation group. The rats in both SPS model group and SPS combined with social isolation group were modeled by single-prolonged stress,and the rats in the SPS combined with social isolation group were raised separately after modelling. The weight gaining,the total movement distance in open field test,the frequency of grid crossing,the single maximum movement distance,and the freezing frequency and time durations in the freezing behavior test were measured after 7 days of modeling. Results Compared with the normal group,the weight gaining and the single maximum movement distance of the rats in the SPS model group were significantly decreased(P < 0.01),as well as the total movement distance and the frequency of grid crossing(P < 0.05),while the freezing frequency and time in the freezing behavior test were significantly increased(P < 0.01). Compared with the normal group,the weight gaining and crossing times of the rats in the SPS combined with social isolation group was decreased(P <0.05),and the freezing frequency and time durations in the freezing behavior test were increased(P < 0.05). In addition, compared with the SPS model group,the total movement distance in the open field test,the frequency of grid crossing and the single maximum movement distance of the rats in the SPS combined with social isolation group were increased(P < 0.05). Conclusions The rat model of post-traumatic stress disorder is successfully established by single-prolonged stress, and 7 days of social isolation may alleviate the anxiety state of SPS model rats.
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Objective:To investigate the role of miR-581 overexpression on the proliferation of human colorectal cancer cell line SW620. Methods:The expression group,colorectal cancer SW620 cells were transfected with recombinant lentivirus vector ( LV-miR-581) and miR-581 mimics(miR-581),the negative control group were transfected with negative control lentiviral vector (LV-GFP) and negative control mimics (vector). The mRNA expression of miR-581 was identified by qRT-PCR. Proliferation of the cells were detected by CCK8 assary and colony forming assary. Results:The expression of miR-581 at mRNA significantly increased in LV-miR-581 group compared with control groups were detected by qRT-PCR ( P<0. 05 ) . Up-regulation of miR-581 markedly enhanced human colorectal cancer SW620 cells proliferation than those in the cells transfected with control vector ( P<0. 05 ) . Conclusion: Forced expression of miR-581 accelerates the proliferation of colorectal cancer SW620 cells.
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Objective:To study the expression and regulation of TLR2/4 in mycobacterium tuberculosis heat shock proteins 16. 3 (mycobacterium tuberculosis heat shock proteins 16. 3,MTB Hsp16. 3) effect on mouse bone marrow-derived macrophages in vitro. Methods:Bone marrow cells were isolated from tibia and femurs of BALB/c mice and incubated with GM-CSF,then detected the expression of CD11b and F4/80 with flow cytometry and observed morphology. The M0 macrophages were stimulated with MTB Hsp16. 3 for 0 h,12 h,24 h,36 h,48 h and 72 h. Real-time PCR detected the expression of TLR2/4 in intracellular at different time point. Silencing macrophages cell surface TLR2/4 molecules by siRNA technology which stimulated with MTB Hsp16. 3 for 0 h,12 h,24 h,36 h,48 h and 72 h. Real-time PCR detected the expression of TLR2/4,Ym-1,Fizz1,IL-10,TNF-α,iNOS and TGF-βin intracellular at different time point. Results:Morphology analysis showed that MTB Hsp16. 3 stimulated macrophages were round cells stretching out pseudopodia,whereas MTB Hsp16. 3 stimulated silencing TLR2/4 macrophages had elongated fibroblastoid. Real time PCR detected the expression of TLR2/4 were upregulated after MTB Hsp16. 3 stimulated M0 macrophages. MTB Hsp16. 3 stimulated silencing TLR2/4 macrophages the expression of IL-6, TNF-α, iNOS were upregulated, whereas IL-10, TGF-β, Ym-1 and Fizz1 were downregulated. Conclusion:MTB Hsp16. 3 may stimulated M0 macrophages to M2 macrophages and suppress M1 macrophages through binding with TLR2/4 receptor,which may be involved the progresss of MTB evaded macrophage phagocytosis.
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Meridians in human body were classified as white meridian and black meridian according to Tibetan medicine.Season and environment,improper diet,toxic heat and trauma were recognized as main reasons damaging the white meridian in Tibetan Medicine,leading to the emerge of white meridian disease induced by Long (one of the three factors) and blood disorder.White meridian disease in Tibetan medicine involved a series diseases,such as many clinical diseases,due to the damage of white meridian system caused by pathogenic factors.Stroke also belonged to white meridian disease.Drugs and treatments were selected based on the nature of disease such as cold and heat,onset,thelocation of disease and the three factors (Chi Ba,Long and Pei Gen).It was the fundamental principle of the treatment rules of white meridian disease in Tibetan medicine,namely,prescribing medication with the rule of diagnosis and treatment,comprehensive analysis of the causes of diseases and mastering the change law of diseases and syndromes in clinic.
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Objective:To investigate the effects of RNA interference(RNAi)-mediated silencing of Peroxiredoxin 1(PRDX1)gene on the invasion and migration of human colorectal cancer SW480 cells.Methods: Lentiviruses negative control vector and PRDX1 RNAi were transfected respectively into colorectal cancer SW480 cells.The transfected cells were divided into PRDX1 silencing group(si-PRDX1)and negative control group(Vector).The expressions of PRDX1 mRNA and protein in SW480 cells were exa mined by quantitative real-time PCR(qRT-PCR)and immunoblotting(Western blot),respectively.The cell migration and invasion capabilities were evaluated with transwell chamber assay and transwell chamber,respectively.The protein expressions of TIMP-2,MMP-2 and MMP-9 were detected by Western blot.Results: Compared with control group,the expressions of PRDX1 mRNA and protein were significantly decreased in PRDX1 silencing group(P<0.01),PRDX1 gene silencing cell line was successfully constructed.The levels of invasion and migration capacities of SW480 cells transfected with si-PRDX1 were lower than those in the cells transfected with control-siRNA(vector)(P<0.01).The expression of TIMP-2 was significantly increased,while the expressions of MMP-2 and MMP-9 were significantly decreased(P<0.05).Conclusion: Silencing of PRDX1 inhibits the invasion,migration and metastasis of human colorectal cancer SW480 cells by regulating the expressions of TIMP-2,MMP-2 and MMP-9.
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Objective:To investigate the effects of Gab2 overexpression on the proliferation and migration of human colorectal cancer cell line SW480.Methods: The experimental group (LV-Gab2-GFP group),colorectal cancer SW480 cells were transfected with recombinant lentivirus vector (LV-Gab2-GFP),the negative control group was transfected with negative control lentiviral vector ( LV-GFP) ,and the blank control group without any treatment.The mRNA and protein expression of Gab 2 in cells were identified by RT-PCR and Western blot respectively.Proliferation of the cells was detected by CCK-8 colorimeter and colony forming assay.Wound-healing assay was used to determine the cells migration .Results: RT-PCR and Western blot demonstrated that Gab 2 mRNA and protein expression significantly increased in LV-Gab2-GFP group compared with control groups;overexpression of Gab2 markedly enhanced human colorectal cancer SW 480 cells proliferation and migration compared with control groups .Conclusion:Overexpression of Gab2 accelerates human colorectal cancer SW 480 cells proliferation and migration.