Expression of HMGA1 in uveal melanoma and its effect on cell proliferation and invasion / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the expression of high mobility group A1(HMGA1)protein in uveal melanoma(UM)tissues, and the effect of inhibiting the expression of <i>HMGA1</i> on cell proliferation and invasion. <p>METHODS: A total of 53 cases(53 eyes)of UM patients who underwent surgical treatment in our hospital from February 2014 to August 2019 were selected. In the same period, 34 cases(34 eyes)of normal uveal tissues removed from the eye due to trauma were selected. The expression of HMGA1 protein in tissues was detected by immunohistochemistry. The human UM cell line M23 was cultured and divided into <i>HMGA1</i> downregulation group, negative control group and blank group, respectively, transfected with <i>HMGA1</i> interference sequence, negative control sequence and without any treatment. The expression of <i>HMGA1 </i>was detected by real-time quantitative PCR. The cell proliferation ability was detected by CCK-8 method, and the cell migration and invasion abilities were detected by Transwell method.<p>RESULTS: The positive expression rate of HMGA1 protein in UM tissue was 77%, which was higher than that in the normal uveal tissue, which was 29%(<i>P</i><0.001). Compared with the no scleral occurring infiltration, no ciliary body involving, and no extraocular growth, the positive expression rates of HMGA1 proteins in the scleral infiltration, ciliary body involving, and extraocular growth occurring tissues were increased(all <i>P</i><0.05). The relative expression level of <i>HMGA1</i> mRNA in cells in the <i>HMGA1</i> downregulation group was lower than that in the negative control group and the blank group. Compared with the negative control group and the blank group, the absorbance <i>OD</i> values of cells in the <i>HMGA1</i> downregulation group at 24, 48, 72 and 96h were decreased(<i>P</i><0.05). The number of migrating cells and the number of invading cells in the <i>HMGA1</i> downregulation group was significantly less than those in the negative control group and the blank group(<i>P</i><0.05). <p>CONCLUSION: The positive expression rate of HMGA1 protein in UM tissue is increased. Down regulation the expression of <i>HMGA1</i> in M23 cells can reduce cell proliferation and inhibit cell migration and invasion.

Expression of protease-activated receptor 2 in uveal melanoma and its effect on cell proliferation and invasion / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the expression of protease-activated receptor 2(PAR2)in uveal melanoma(UM), and the effects of silencing the expression of PAR2 gene on proliferation and invasion of human UM cell line M23. <p>METHODS: A total of 45 patients(45 eyes)with UM who underwent surgical treatment with complete information in our hospital were selected from February 2012 to December 2017. In the same period, 30 patients(30 eyes)who underwent eyeball removal due to ocular trauma and most of the uvea were normal were selected. Real-time quantitative PCR was used to detect the expressions of PAR2 gene in UM and normal choroidal tissues. M23 cells were cultured and divided into PAR2 interference group, negative control sequence group and blank group. Real-time quantitative PCR was used to detect the expression of PAR2 gene in cells. MTT assay was used to detect cell proliferation, and transwell assay was used to detect cell migration and invasion. <p>RESULTS: The relative expression level of PAR2 mRNA was 1.73±0.13 in UM tissues, and 1.06±0.10 in normal choroid tissues(<i>t</i>=23.732, <i>P</i><0.001). The relative expression level of PAR2 mRNA in UM tissues was associated with pathological type, scleral invasion, optic disc involvement and extraocular growth(<i>P</i><0.05). The relative expression level of PAR2 mRNA in PAR2 interference group was lower than that in negative control sequence group and blank group(<i>P</i><0.05). The absorbance <i>A</i> values at 24h, 48h, 72h and 96h in the PAR2 interference group cells were lower than those in negative control sequence group and blank group(<i>P</i><0.05). The number of migrated cells and the number of invasive cells in PAR2 interference group were lower than those in negative control sequence group and blank group(<i>P</i><0.05). <p>CONCLUSION: PAR2 was highly expressed in UM tissues and was associated with high risk of tumor metastasis. Specific silencing of PAR2 gene expression in M23 cells could effectively inhibit cell proliferation, migration and invasion.

Lithium chloride promotes neuronal differentiation of rat bone marrow mesenchymal stem cells by modulating autophagy / 中国病理生理杂志

AIM:To study the influence of lithium chloride (LiCl) on the neuronal differentiation of rat bone marrow mesenchymal stem cells (MSCs), and to explore whether autophagy was involved in this process .METHODS:MSCs were isolated and cultured in vitro.The cells were divided into LiCl group and control group .MSCs were treated withβ-mercaptoethanol as an inducer for triggering the cells to differentiate into neurons .The expression of neuronal markers-neuron specific enolase (NSE) and microtubule-associated protein-2 (MAP-2), and autophagic marker-microtubule-associ-ated protein 1 light chain 3 ( LC3) were measured by immunofluorescence method and Western blot .An autophagy activator rapamycin and autophagy inhibitor 3-methyladenine (3-MA) were applied to modulate the autophagy in the LiCl treated-cells.The protein expression of NSE and MAP-2 were determined by Western blot .RESULTS: After induction, the ex-pression of NSE and MAP-2 were increased .The percentage of NSE-and MAP-2-positive cells and the expression of NSE and MAP-2 in the LiCl group were greater than those in control group (P<0.05).After induction, the number of LC3-positive dots and the expression of LC3-Ⅱin LiCl group were greater than those in control group (P<0.05).The expres-sion of NSE and MAP-2 increased when the autophagy was modulated by rapamycin in LiCl treated -cells, and on the contra-ry, the expression of NSE and MAP-2 were inhibited as autophagy was modulated by 3-MA.CONCLUSION: Lithium chloride may promote the neuronal differentiation of rat bone marrow mesenchymal stem cells by modulating autophagy .

Impact of shengmal injection on changes of immunological function in patients after cardiopulmonary bypass / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the impact and mechanism of Shengmai Injection (SMI) on the immunological function changes after cardiopulmonary bypass.</p><p><b>METHODS</b>Forty patients with rheumatic heart valve disease were selected and assigned randomly to two groups: 20 in the control group and 20 in the SMI group. Peripheral blood samples were taken at various time points, i.e. 3 days before operation (T1), 10 min after terminal of CPB (T2), the first (T3), third (T4), and seventh (T5) day after operation, for counting white blood cell (WBC), neutrophils and lymphocytes; percentage of T lymphocytes (CD3+ mononuclear cells) and its subsets (CD4+ and CD8+) to calculate CD4+/CD8+ ratio; and the serum content of immunoglobulins (IgG, IgA, IgM) as well as serum concentration of interleukin-8 (IL-8) and interleukin-10 (IL-10) were assayed.</p><p><b>RESULTS</b>Compared with the control group, in the SMI group, WBC and neutrophil count were lower at T2 (P < 0.01); percentages of CD3+ and CD4+ lower at T4 and T5 (P < 0.05 or P < 0.01); percentage of CD8+ higher at T2 to T5 (P < 0.05 or P < 0.01); CD4+/CD8+ ratio lower at T3 to T5 (P < 0.05 or P < 0.01); IgG lower at T2 (P < 0.05); IgA higher at T3 (P < 0.05); IgM higher at T3 to T5 (P < 0.05); IL-8 lower at T2 to T4 (P < 0.05); and IL-10 higher at T2 (P < 0.05).</p><p><b>CONCLUSION</b>Application of SMI in the perioperative period can enhance the humoral immunity and inhibit the cellular immunity after CPB, it could also reduce the systemic inflammatory reaction and improve the prognosis of patients.</p>

The role of p38 MAPK in gastrin-induced u-PA expression in human colon cancer cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effect of gastrin on the mRNA and protein expression of urokinase-type plasminogen activator (u-PA) in human colon cancer cells and detect the role of p38 MAPK in this process.</p><p><b>METHODS</b>Lipofectin method was used to transfect pCR3. 1/CCK2R vector expressing gastrin receptor into a colon cancer cell line colo320. Gastrin and gastrin antagonist were used to up-regulate and down-regulate the signaling pathway, respectively. Human colon cancer colo320 cells and colo320/ CCK2,R cells were cultured and then stimulated with gastrin for different time; SB203580 was added into culture medium to prevent p38 kinase pathway before incubating with gastrin; Western blot and RT-PCR were used to examine the u-PA expression. Western blot was employed to detect p38 kinase phosphorylation.</p><p><b>RESULTS</b>Gastrin increased evidently the mRNA and protein expressions of u-PA and induced p38 kinase phosphorylation in colo320/CCK,R cells time-dependently. However, the extent of enhancement of u-PA and p38 MAPK expression in colo320 cells was much less than that in colo320/CCK2R cells. The gastrin antagonist L-365, 260 showed an effect of competitive inhibition on gastrin-induced u-PA expression and p38 kinase phosphorylation. The inhibitor SB203580 could sufficiently suppress gastrin-induced p38 kinase phosphorylation and significantly attenuate gastrin-induced u-PA mRNA and protein expressions in colo320/ CCK2 R cells in a dose-dependent manner.</p><p><b>CONCLUSION</b>Gastrin-gastrin receptor signal transduction pathway can obviously induce u-PA expression in human colon cancer cells via activating the phosphorylation of p38 kinase.</p>

Immunohistochemical observation of heart-type fatty acid binding-protein in early myocardial ischemia / 法医学杂志

OBJECTIVE@#The sensitivity of heart-type fatty acid binding-protein (H-FABP) in the postmortem diagnosis of myocardial ischemia was explored.@*METHODS@#The changes of H-FABP staining in normal, infarcted and suspected ischemia of myocardial cells were studied by immunohistochemistry.@*RESULTS@#There was no depletion in normal control group, and obvious depletion was observed in myocardial infarcted group. Among 9 suspected myocardial ischemia group, 3 cases showed obvious depletion and 3 cases showed vague depletion for H-FABP, there were obvious depletion of Mb in 4 suspected myocardial ischemia cases and vague depletion in 2 cases for Mb. It is indicated that H-FABP can be used to diagnose early myocardial ischemia.@*CONCLUSION@#H-FABP is quite sensitive and useful for the diagnosis of early myocardial ischemia.