Objective To study the mechanisms of Re Bi granule on treating RA.Methods Make SD maleness rats Adjuvant-Induced Arthritis(AA)which was derived by Freund's complete adjuvant(CFA)as investigation object.Make Wang Bi granule as positive control medicine,and divide it into six groups(blank group,model group,control group,Infusion of Re Bi granule high dose group and midst dose group and low dose group).Observe Re Bi granule's influence and mechanism to synovial cell apoptosis and T cell subsets.Results Re Bi granule could distinctly heighten synovial cell apoptosis ratio of AA rats,heighten CD8+cell's number and reduce CD4+cell's number,lower CD4+/CD8+ ratio,adjust T cells for two-way.Conclusion Re Bi granule not only is an effective treatment way for rheumatoid arthritis,but also can improve the abnormal immune status.

Objective To construct a new recombinant immunotoxin expression vector by using human VEGF165 and a truncated pseudomonas exotoxin A ramification (PE38) gene, and explore the expression of the VEGF165-PE38 fusion protein in HEK293 cells. Methods VEGF165 was cloned by polymerase chain reaction (PCR). PE38 gene was gained from an vector plasmid pRB391 by restriction endonuclease digestion, and then inserted to the eukaryotic expression vector pIRES2-EGFP. After the eukaryotic recombinant vector pIRES2-VEGF165-PE38-EGFP was identified by restriction endonuclease digestion and sequence analysis, the vector was transfected into HEK293 cells by liposome protocol. RT-PCR and ELISA method was used to confirm the expression of the fusion gene in the HEK293 cells. Results Restriction endonuclease digestion and sequence analysis revealed the VEGF165-PE38 fusion gene was cloned into the eukaryotic expression plasmid vector pIRES2-EGFP successfully. The pIRES2-VEGF165-PE38-EGFP fusion gene could express in the HEK293 cells. Conclusion The result provide the basis for search of the targeted cytotoxic activity to tumor vascular endothelial cells and may have some potential value in clinical application.

OBJECTIVETo investigate the effect of liraglutide, an analogue of glucagon-like peptide-1, on the proliferation and migration of cardiac microvascular endothelial cells (CMECs) and explore the mechanism.

METHODSIn vitro cultured CMECs of SD rats were purified by differential adhesion method and identified immunocytochemically using CD31 antibody and factor VIII. MTT assay was performed to assess the proliferation of the first-generation cells exposed to different concentrations (0-1000 nm/L) of liraglutide. Western blotting was used to detect the activation of PI3K/Akt and MAPK/ERK signaling pathways. BrdU fluorescent labeling and scratch assay were performed to observe the proliferation and migration of CMECs following liraglutide treatment, and PI3K/Akt and MAPK/ERK pathway inhibitors LY294002 and PD98059, respectively, were used to further confirm the role of these signaling pathways in regulating the proliferation and migration of CMECs.

RESULTSImmunocytochemical staining demonstrated a proportion of double positive cells exceeding 95%. The cells exhibited a logarithmic growth 48 h after plating. Liraglutide exposure concentration-dependently promoted the proliferation of CMECs with the optimal concentration of 100 nmol/L (P<0.05). Liraglutide exposure of the cells for 24 h significantly increased the levels of intracellular phosphorylated Akt and ERK (P<0.05), but pretreatment of the cells with Akt and ERK signaling pathway inhibitors 1 h before liraglutide obviously reversed such effect (P<0.05). BrdU and scratch assay showed that 100 nmol/L liraglutide significantly promoted the proliferation and migration of CMECs (P<0.05), but such effects were obviously suppressed by Akt and ERK inhibitors (P<0.05).

CONCLUSIONLiraglutide promotes the proliferation and migration of CMECs in vitro via PI3K/Akt and MAPK/ERK signaling pathways.

Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chromones ; Endothelial Cells ; cytology ; drug effects ; Flavonoids ; Glucagon-Like Peptide 1 ; analogs & derivatives ; pharmacology ; Liraglutide ; MAP Kinase Signaling System ; Morpholines ; Myocardium ; cytology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effect of liraglutide, an analogue of glucagon-like peptide-1, on the proliferation and migration of cardiac microvascular endothelial cells (CMECs) and explore the mechanism. Methods In vitro cultured CMECs of SD rats were purified by differential adhesion method and identified immunocytochemically using CD31 antibody and factor VIII. MTT assay was performed to assess the proliferation of the first- generation cells exposed to different concentrations (0-1000 nm/L) of liraglutide. Western blotting was used to detect the activation of PI3K/Akt and MAPK/ERK signaling pathways. BrdU fluorescent labeling and scratch assay were performed to observe the proliferation and migration of CMECs following liraglutide treatment, and PI3K/Akt and MAPK/ERK pathway inhibitors LY294002 and PD98059, respectively, were used to further confirm the role of these signaling pathways in regulating the proliferation and migration of CMECs. Results Immunocytochemical staining demonstrated a proportion of double positive cells exceeding 95%. The cells exhibited a logarithmic growth 48 h after plating. Liraglutide exposure concentration-dependently promoted the proliferation of CMECs with the optimal concentration of 100 nmol/L (P<0.05). Liraglutide exposure of the cells for 24 h significantly increased the levels of intracellular phosphorylated Akt and ERK (P<0.05), but pretreatment of the cells with Akt and ERK signaling pathway inhibitors 1 h before liraglutide obviously reversed such effect (P<0.05). BrdU and scratch assay showed that 100 nmol/L liraglutide significantly promoted the proliferation and migration of CMECs (P<0.05), but such effects were obviously suppressed by Akt and ERK inhibitors (P<0.05). Conclusion Liraglutide promotes the proliferation and migration of CMECs in vitro via PI3K/Akt and MAPK/ERK signaling pathways.

Objective To investigate the effect of liraglutide, an analogue of glucagon-like peptide-1, on the proliferation and migration of cardiac microvascular endothelial cells (CMECs) and explore the mechanism. Methods In vitro cultured CMECs of SD rats were purified by differential adhesion method and identified immunocytochemically using CD31 antibody and factor VIII. MTT assay was performed to assess the proliferation of the first- generation cells exposed to different concentrations (0-1000 nm/L) of liraglutide. Western blotting was used to detect the activation of PI3K/Akt and MAPK/ERK signaling pathways. BrdU fluorescent labeling and scratch assay were performed to observe the proliferation and migration of CMECs following liraglutide treatment, and PI3K/Akt and MAPK/ERK pathway inhibitors LY294002 and PD98059, respectively, were used to further confirm the role of these signaling pathways in regulating the proliferation and migration of CMECs. Results Immunocytochemical staining demonstrated a proportion of double positive cells exceeding 95%. The cells exhibited a logarithmic growth 48 h after plating. Liraglutide exposure concentration-dependently promoted the proliferation of CMECs with the optimal concentration of 100 nmol/L (P<0.05). Liraglutide exposure of the cells for 24 h significantly increased the levels of intracellular phosphorylated Akt and ERK (P<0.05), but pretreatment of the cells with Akt and ERK signaling pathway inhibitors 1 h before liraglutide obviously reversed such effect (P<0.05). BrdU and scratch assay showed that 100 nmol/L liraglutide significantly promoted the proliferation and migration of CMECs (P<0.05), but such effects were obviously suppressed by Akt and ERK inhibitors (P<0.05). Conclusion Liraglutide promotes the proliferation and migration of CMECs in vitro via PI3K/Akt and MAPK/ERK signaling pathways.

Objective To determine the impact of family cohesion and adaptability on depression of young patients with stroke.Methods A total of 188 young patients with stroke were recruited from rehabilitation departments of three class Ⅲ grade A hospitals in Wenzhou by convenience sampling method from January to December 2016. The status quo of family cohesion and adaptability and depression of these patients was investigated, and the relationship between them was analyzed by correlation and regression analysis.Results The total score of depression in young patients with stroke was (25.71±6.11)points. The correlative analysis showed that there were negative correlations between score of actual family cohesion, score of actual family adaptability and score of depression(P<0.05). The regression analysis showed that actual family cohesion and actual family adaptability could independently explain 41.6% of the variance for depression of young patients with stroke after controlling demographic variables.Conclusions Family cohesion and adaptability could predict depression of young patients with stroke. Improve the actual family cohesion and actual family adaptability can help to reduce the level of depression in young stroke patients

Aim To investigate the effects of Gubu decoction combined with intraluminal reconstruction on bi-pedal vibration sensation threshold and microcirculation in patients with lower extremity arteriosclerotic occlusive disease(LEAOD).Methods A total of 152 patients who came to Lishui Gentral Hospital for LEAOD treatment from April 2021 to April 2023 were divided into the observation group(intraluminal reconstruction+Gubu decoction,76 cases)and the control group(intraluminal reconstruction,76 cases)by random number table method,and the traditional Chinese medicine(TCM)syndrome scores,clinical efficacy,microcirculation indicators and bifoot vibration sensation threshold of the two groups were compared.Results After treatment,the scores of TCM syndromes such as limb pain,limb numbness,intermittent claudication,and limb fear of cold decreased after treatment in both groups,and the decrease of ob-servation group was more obvious(P＜0.05).The clinical effect of observation group(96.05％)was significantly higher than that of control group(84.21％)(P＜0.05).After treatment,the flow state integral,tube loop integral and periloop integral decreased,and the apical vessel diameter increased in both groups,and the changes of various indexes in the ob-servation group were more significant(P＜0.05).After treatment,the threshold of sensation in both groups decreased,and the observation group was significantly lower(P＜0.05).Conclusion Gubu decoction combined with intraluminal reconstruction can effectively reduce the foot sensation threshold and improve the microcirculation of affected limbs in LEAOD patients.