Objective: To investigate the relationship between blood level of Galectin-3 and the severity of coronary lesions in patients with coronary artery disease (CAD). Methods: A total of 158 consecutive subjects received coronary artery angiography (CAG) in Yantai Yu huangding hospital were studied and they were divided into 2 groups: Control group,n=38 individuals with normal coronary artery, CAD group,n=120 patients with at least 1 coronary branch stenosis ≥ 50%. CAD group was further divided into 2 sets of subgroups:①By the number of branches involved, as Single vessel disease, n=42, Double vessels disease including LM,n=40 and Triple vessel disease,n=38.②By the quartile of Gensini score as 1st quartile group, the patients with Gensini score ≤ 18.5, 2nd quartile group,18.5 < Gensini score ≤ 45.0, 3rd quartile group, 45.0 < Gensini score ≤ 71.5 and 4th quartile group, Gensini score>71.5,n=30 in each subgroup. Blood levels of Galectin-3 were examined, and the severity of coronary lesions was evaluated by both branch numbers and Gensini score analysis. Results: Compared with Control group, the blood level of Galectin-3 was higher in CAD group,10.66 (5.81, 16.17) ng/ml vs 18.3(1.14, 2.52) ng/mlP<0.01; with the more branches of coronary lesions involved, the blood levels of Galectin-3 increased accordingly, all P <0.01. With the elevation of Gensini score, the levels of Galectin-3increased accordingly, except for the difference between the 3rd quartile group and 4th quartile group, all P <0.01.With adjusted other factors, blood levels of Galectin-3 were positively related to the number of coronary branchlesions (r =0.52, P <0.01) and Gensini score levels (r =0.17, P =0.04).Conclusion: Blood level of Galectin-3 is positively related to the severity of coronary lesions whichimplies that Galectin-3 may have potential detrimental effect on the occurrence and development of coronaryatherosclerosis.

Objective To obtain highly purified oligodendrocyte precursor lineage cells in vitro and make identification.Methods The oligodendrocyte precursors were separated from astrocyte by orbital shaker and further purified by differential adhesion,and finally cultured in chemically defined serum-free medium,with appended neurotrophin 2(N2),platelet-derived growth factor(PDGF),basic fibroblast growth factor(bFGF).Immunofluorescence assay was applied to identify the separated cells with A2B5,O4,O1 and glial fibrillary acidic protein(GFAP) antibodies.Results Over 95% of cultured oligodendrocyte precursor cells were obtained.The oligodendrocyte progenitors were A2B5 and O4 positive,immature oligodendrocytes were O4 and O1 positive while GFAP were negative.Conclusions Separation and purification by shaking and differential adhesion and chemically defined medium are suitable and effective to obtain highly purified oligodendrocyte precursor cells.Cell output will increase notabily and rest in immature phase by appending both N2,PDGF and bFGF.

Objective: To explore the protective roll of ifbroblast growth factor 21 (FGF21) in endoplasmic reticulum stress (ERS) induced rat's H9c2 cardiomyocyte apoptosis with its mechanism. Methods: pcDNA4 was used as gene vector, pcDNA4-FGF21 plasmid was constructed and transfected into rat's H9c2 myocardiocytes for 48 h. ERS model was established by 10 μM tunicamycin (TM) induction for 24 h. The experiment was conducted in 4 groups:①Control group,②TM group, the cells were treated by TM,③pcDNA4-FGF21+TM group,④pcDNA4+TM group. The expressions of FGF21, protein kinase R-like ER kinase (PERK) and c-Jun N-terminal kinases (JNK) mediated apoptosis pathway related protein were measured by Western blot analysis; cell survival rate was examined by CCK-8 method and apoptosis rate was detected by TUNEL technique. Results: pcDNA4-FGF21 vector was successfully constructed and overexpressed in H9c2 myocardiocytes. Compared with Control group, TM group and pcDNA4+TM group had up-regulated endogenous FGF21 expression, increased PERK and JNK mediated apoptosis pathway related protein expression; reduced cell survival rate and elevated apoptosis rate. Compared with TM group and pcDNA4+TM group, pcDNA4-FGF21+TM group had down-regulated PERK and JNK mediated apoptosis pathway related protein expression; increased cell survival rate and decreased apoptosis rate. Conclusion: FGF21 overexpression can reduce ERS induced apoptosis rat's H9c2 myocardiocytes which might be partly related for inhibiting PERK and JNK mediated signal transduction of apoptosis pathway.

Humans ; Integrative Medicine ; Medicine, Chinese Traditional ; trends

Objective To observe the effects of hyperbaric oxygen combined with rehabilitation training on the motor ability of ischemic stroke patients.Methods Eighty ischemic stroke patients were randomly divided into a treatment group and a control group,with 40 cases in each group.The control group was treated with Bobath' s approach,while the treatment group was treated with Bobath's methods supplemented by hyperbaric oxygen therapy.The United States National Institutes of Health Stroke Scale (NIHSS),the Fugl-Meyer motor assessment scale (FMA) (lower part) and each patient's maximum walking speed (MWS) were used to evaluate the patients before,and after 8 weeks of treatment.Results After treatment the average NIHSS score,FMA score and MWS were 4.17 ± 1.4％,31.2 ± 3.3 and 54,.3 ± 16.2 m/min,respectively,in the treatment group.The control group' s results were 6.81 ± 1.2％,26.2 ± 2.2 and 45.6 ± 18.3 m/min.The intra-group differences in evaluation results before and after treatment were statistically significant in both groups.An inter-group comparison showed that the treatment group performed significantly better after treatment than the control group in terms of FMA and MWS.After treatment,the treatment group showed significantly better walking performance in terms of cadence,stride length,step length on the affected side,gait cycle and double support duration.Conclusion Hyperbaric oxygen can make rehabilitation training more effective in improving the neurologic deficits,motor function and walking ability of hemiplegic stroke survivors.

BACKGROUND: Matrix metalloproteinase (MMP) is a kind of protease family, its activity can be inhibited by tissue inhibitor of metalloproteinase (TIMP), especially by TIMP-3.OBJECTIVE: To fully isolate and purify natural TIMP-3, and to create enzyme-linked immunoassay of TIMP-3.DESIGN: Single-sample observation SETTING: Central Laboratory of Shenyang Medical College MATERIALS: Human placenta from Department of Obstetrics & Gynecology, Fengtian Hospital Affiliated to Shenyang Medical College (Informed consent was obtained from the relatives of patients) and MMP-1 from Research Institute. Fuji Chemical Industries, Ltd.METHODS: This experiment was conducted in the Central Laboratory of Shenyang Medical College between March 2001 and May 2002. Firstly,4 mol/L urea Tris-buffer solution (pH8.0) was used to prepare homogenate solution of placenta. Secondly, homogenate solution was performed chromatography through CM52 positive ion-exchange resin and Sephacryl S200 gel filtration. Thirdly, relative molecular weight and purity were detected by SDS- polyacrylamidedel gel electrophoresis (SDS-PAGE).Fourthly, Western blotting was used to identify the characters of purified protein. Fifthly, the inhibitory rate of TIMP-3 to MMP-1 was measured with immumofluorescence method.MAIN OUTCOME MEASURES: ① The relative molecular mass of protein showed by PAGE. ② Results of Western blot. ③ The inhibitory rate of TIMP-3 for MMP-1. ④ The recovery rate of TIMP-3 following purification.RESULTS: ①The isolated and purified TIMP-3 from human placenta consisted of non-glycosylated and glycosylated protein, with relative molecular mass of 24 000 and 27 000, respectively. ② The inhibitory concentration of non-glycosylated and glycosylated TIMP-3 was 1.1×1010 mol/L and 1.2×1010 mol/L respectively to MMP-1. The inhibitory concentration of placenta-derived TIMP-3 was significantly higher than that of recombinant TIMP-3 for MMP-1. ③ The recovery rate of TIMP-3 was 23.4% following two-step chromatography.CONCLUSION: Extracellular matrix of human placenta-derived TIMP-3 consists of non- glycosylated protein and glycosylated protein; Two kinds of purified TIMPs-3 have remarkable inhibitory concentration for MMP-1, and significantly higher in comparison with recombinant metalloproteinase inhibitory factor-3.

Female ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; Retrospective Studies

Objective To observe the effect of adenosine monophosphate activated protein kinase (AMPK) on attenuating inflammation in fibrosis induced hy acute ischemia reperfusion injury (IRI) in mice.Methods Forty eight male C57BL/6 mice were randomly divided into four groups:sham operation group (sham group),IRI group,AMPK inhibitor+IRI group (AMPK/IRI group) and normal saline+IRI group (NS/IRI group),12 mice each group.The mice with renal IRI were occluded for 30 min through clipping bilateral renal pedicle,then released renal perfusion.Mice in sham group were performed the separation of renal pedicle without clipping.Mice in AMPK/IRI group and NS/IRI group were respectively intraperitoneal injected AMPK inhibitor and normal saline before IRI.At the 2 d after operation,6 randomly-selected mice from each group were blooded by extraction eyeball to detect BUN and Scr.The renal histopathological changes were observed through HE staining.The mRNA expression of IL-1β,IL-6 and TNF-α was detected by real time PCR,and the level of AMPK phosphorylation was detected by Western blotting.At the 14 d after operation,Collagen 1 (COL1),α-SMA and fibronectin (FN) were detected by immunofluorescence and Western blotting in 6 remained mice from each group.The degree of kidney fibrosis was observed through sirus red staining.Results Compared with those in sham group,tubular interstitial damage was aggravated (P ＜ 0.05),BUN and Scr were increased (P ＜ 0.05),the mRNA expression of IL-1β,IL-6 and TNF-α was increased at the 2 d after operation (all P ＜ 0.05),and the level of AMPK phosphorylation was activated in IRI group and NS/IRI group (all P ＜ 0.05);the degree of kidney fibrosis and the expression of COL1,α-SMA and FN were increased obviously at the 14 d (all P ＜ 0.05).Compared with those in IRI group,in AMPK/IRI group tubular interstitial damage was aggravated (P ＜ 0.05),BUN and Scr were increased (all P ＜ 0.05),the mRNA expression of IL-1β,IL-6 and TNF-α was increased at the 2 d (all P ＜ 0.05),and the level of AMPK phosphorylation was decreased (P ＜ 0.05).Moreover,the degree of kidney fibrosis and the expression of COLI,α-SMA and FN were increased obviously at the 14 d in AMPK/IRI group (all P ＜0.05).Conclusions AMPK can ameliorate the acute renal ischemia reperfusion injury induce fibrosis in mice,and the mechanism may be related to the decrease of inflammatory reaction.

AIM:To explore the effects of galectin-3 (GAL-3) on the viability, migration and inflammation of human umbilical vein endothelial cells (HUVECs) and the mechanisms.METHODS:The HUVECs were cultured in vitro and treated with GAL-3 recombinant protein at 2 mg/L or GAL-3 short hairpin RNA (shRNA).The HUVECs were divided into normal group, recombinant GAL-3 group, shControl group and GAL-3-shRNA group.The mRNA expression of GAL-3, monocyte chemotactic protein (MCP)-1, IL-6, matrix metalloproteinase (MMP)-9 and cyclin D1 was detected by real-time quantitative PCR,and the protein expression of GAL-3, IL-6 and MCP-1 was detected by Western blot.The secretion levels of MCP-1 and IL-6 in the culture medium were measured by ELISA.The viability and the ability of migration of the HUVECs were examined by CCK-8 assay and wound healing assay.The protein levels of heat shock protein 90 (HSP90), ERK1/2, p-ERK1/2, JNK and p-JNK were determined by Western blot.RESULTS:The expression of GAL-3, MCP-1 and IL-6 at mRNA and protein levels, the mRNA expression of MMP-9 and cyclin D1, and the secretion levels of MCP-1 and IL-6 in the culture medium were significantly higher than those in normal group (P<0.05) after the HUVECs were treated with GAL-3 recombinant protein.However, these molecules mentioned above in GAL-3-shRNA group were significantly lower than those in normal group and negative control group (P<0.05).Compared with normal group, the viability and migration ability of the HUVECs in recombinant GAL-3 group were significantly increased, but the viability and migration ability of the HUVECs in GAL-3-shRNA group were lower than those in normal group and shControl group (P<0.05).In addition, the protein levels of p-ERK1/2 and HSP90 in recombinant GAL-3 group were higher than those in normal group (P<0.05), but those in GAL-3-shRNA group were lower than those in normal group and shControl group (P<0.05).The protein level of p-JNK was not oviously changed among the 4 groups.CONCLUSION:GAL-3 is involved in regulating the cell growth, migration and the release of inflammatory cytokines in vascular endothelial cells, which may be mediated by HSP90-ERK1/2 signaling pathway.

The pollution hazard of formaldehyde in low level from indoor air is worldwide.Many researches have been focused on the ornamental plant purification for indoor air formaldehyde in low concentration.Although a numbers of literatures have documented that gaseous formaldehyde can be removed by many plant species,a long term purification efficiency of the plants for formaldehyde is uncertain.The research in the future works should be focused on phytotoxieity of formaldehyde,a long term purification efficiency of plants,soil contribution to formaldehyde removal under plant-soil system,effects of complex pollutants on the purification of plants for formaldehyde,and volatile organic compounds from the plants which can remove formaldehyde.