Objective To analyze the clinical application value of serum homocysteine(Hcy) detection in diagnosing acute myo-cardial infarction .Methods 78 cases of acute myocardial infarction in the hospital from January to December 2013 were selected as the acute myocardial infarction group ,69 cases of unstable angina as the unstable angina group and contemporaneous 78 healthy per-sons undergoing the physical examination as the control group .The serum Hcy ,myoglobin and creatine kinase isoenzyme were de-tected and the detection results were performed the statistical analysis .Results The serum Hcy levels and the positive rate in the a-cute myocardial infarction group were significantly higher than those in the unstable angina group (P<0 .05) ,but serum myoglobin and creatine kinase isoenzyme had no statistical differences in the concentration and positive rate between these two groups (P>0 .05) .The serum Hcy concentration and the positive rate in the acute myocardial infarction group and the unstable angina group were higher than those in the control group(P<0 .05) .The ROC curve analysis showed that the efficiency for diagnosing acute my-ocardial infarction from high to low in turn was MYO ,Hcy and CK-MB .Conclusion Serum Hcy may be used as a routine index for diagnosing acute myocardial infarction ,which has certain clinical value for the condition monitoring and prognosis of the disease ,and also has certain clinical significance for the differential diagnosis between acute myocardial infarction and unstable angina .

Animals ; Diabetes Mellitus, Type 2 ; Diabetic Nephropathies ; Disease Models, Animal ; Humans ; Mice ; Rats

BACKGROUNDThe renoprotective mechanisms of adenosine monophosphate (AMP)-activated protein kinase (AMPK) agonist - metformin have not been stated clearly. We hypothesized that metformin may ameliorate inflammation via AMPK interaction with critical inflammatory cytokines. The aim of this study was to observe the effects of metformin on expression of nuclear factor-κB (NF-κB), monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1) and transforming growth factor-beta 1 (TGF-β1) induced by high glucose (HG) in cultured rat glomerular mesangial cells (MCs).

METHODSMCs were cultured in the medium with normal concentration glucose (group NG, 5.6 mmol/L), high concentration glucose (group HG, 25 mmol/L) and different concentrations of metformin (group M1, M2, M3). After 48-hour exposure, the supernatants and MCs were collected. The expression of NF-κB, MCP-1, ICAM-1, and TGF-β1 mRNA was analyzed by real time polymerase chain reaction. Western blotting was used to detect the expression of AMPK, phospho-Thr-172 AMPK (p-AMPK), NF-κB p65, MCP-1, ICAM-1, and TGF-β1 protein.

RESULTSAfter stimulated by HG, the expression of NF-κB, MCP-1, ICAM-1, TGF-β1 mRNA and protein of MCs in group HG increased significantly compared with group NG (P < 0.05). Both genes and protein expression of NF-κB, MCP-1, ICAM-1, TGF-β1 of MCs induced by high glucose were markedly reduced after metformin treatment in a dose-dependent manner (P < 0.05). The expression of p-AMPK increased with the rising of metformin concentration, presenting the opposite trend, while the level of total-AMPK protein was unchanged with exposure to HG or metformin. Conlusion Metformin can suppress the expression of NF-κB, MCP-1, ICAM-1 and TGF-β1 of glomerular MCs induced by high glucose via AMPK activation, which may partly contribute to its reno-protection.

AMP-Activated Protein Kinases ; metabolism ; Animals ; Cells, Cultured ; Glomerular Mesangium ; cytology ; Glucose ; pharmacology ; Mesangial Cells ; drug effects ; metabolism ; Metformin ; pharmacology ; NF-kappa B ; metabolism ; Rats

【Objective】To investigate the mechanism of action of long non-coding RNA highly up-regulated in liver cancer（LncRNA HULC）on the growth of glioblastoma U87 cells in vitro and in vivo.【Methods】The cultured glioblastoma U87 cells were divided into four groups：overexpression group（HULC-over）and its vector control group（VEC），silent expression group（HULC- siRNA）and its negative control group（NC）.Quantitative real- time polymerase chain reaction PCR（qRT-PCR）was used to verify the expression levels of HULC. CCK8 proliferation assay and colony formation assay were adopted to monitor the proliferation of glioblastoma U87 cells. Flow cytometry was utilized to detect the apoptosis of glioblastoma U87 cells. By injecting U87 cells，we divided the orthotopic xenograft mouse model into HULC- over group（n=10），VEC group（n=10），HULC-siRNA group（n=10）and NC group（n=10）accordingly. The survival of the mice in each group was observed. The expression of Ki67 was analyzed by immunohistochemistry. 【Results】 The expression level of HULC was significantly higher in HULC-over group than that in VEC group and significantly lower in HULC-siR NA group than that in NC group（P < 0.01）. The cell proliferation ability was significantly increased in HULC-over group compared with that in VEC group and significantly decreased in HULC- siRNA group compared with that in NC group（P < 0.01 on days2，3and4）. The colony formation rates in VEC group，HULC-over group，NC group and HULC-siRNA group were，respectively，（34.47 ± 1.56）% ，（95.4 ± 2.74）% ，（23.83 ± 0.92）% and （10.23 ± 0.61）% ，which revealed that overexpression of HULC elevated the colony formation rate and silencing expression of HULC reduced the colony formation rate（P < 0.01）. The early apoptosis rates in VEC group，HULC- over group，NC group and HULC- siRNA group were，respectively，（3.55±0.56）% ，（0.09±0.01）% ，（2.89±0.67）% ，and（7.13±0.14）% ，which showed that overexpression of HULC elevated the early apoptosis rate and silencing expression of HULC reduced the early apoptosis rate （P <0.01）. The survival curve of nude mouse indicated shorter survival time in HULC-over group than that in VEC group and longer survival time in HULC-siRNA group than that in NC group（P < 0.05）. Ki67 protein expression was up-regulated in the HULC-over group compared with that in VEC group and down-regulated in the HULC-siRNA group compared with that in NC group（P < 0.05）.【Conclusion】LncRNA HULC can enhance the growth of glioblastoma U87 cells in vitro and in vivo.

Objectives To describe the Epstein Barr virus (EBV) infection of children in Hefei, analyze its epidemiological characteristics, and explore the factors affecting EBV infection. Methods The children as the outpatient in the department of our hospital were recruited as the research subjects from June 2018 to June 2021. Epidemiological data of the research subjects were collected from medical records and the laboratory tests were performed to detect the related serological indicators of EBV. The distribution characteristics of different serological antibodies of EBV were described. According to the types of serological antibodies, all research subjects were divided into three states: primary infection, previous infection and non-infection. Logistics regression was used to analyze the factors affecting the infection status. Results There were 480 children in this study. The mean age and body mass index of all research subjects were 8.7±1.5 years and 20.78±3.2kg/m2, respectively. There were 276 boys(57.50%) and 204 girls(42.50%). 67 children(12.92%) were positive for VCA-IgM antibody. 326 children(67.92%) were positive for VCA-IgG antibody. 290 children(60.42%) were positive for NA-IgG antibody and 25 children(5.21%) were positive for EA-IgG antibody. There was no significant distribution difference of serological antibodies in gender, season of onset, disease duration, fever, angina and enlargement of lymph nodes. However, there were significant distribution differences of serological antibodies among different body mass index(χ²=50.207, P<0.001) and age(χ²=48.295, P<0.001). According to the types of serological antibodies, all research subjects were divided into three states. There were 78(16.25%) children with primary infection, 335(69.79%) with previous infection and 67(13.96%) with non-infection. Only age was the main factor affecting infection status by Logistic regression analysis(P_{primary infection vs. non-infection}<0.001, OR_{primary infection vs. non-infection}=0.580; P_{previous infection vs. non-infection}=0.038, OR_{previous infection vs. non-infection}=2.347). Conclusion The previous infection by EBV is the mainly infection type in children aged 6 to 12 year. The positive VCA-IgG antibody accounts for the most in previous infection. Age is the important influencing factor on EB infection. The younger the age, the higher the probability of primary infection. Besides, the positive VCA-IgM antibody is the main pattern of primary infection in children.

The purpose of this study is to demonstrate if Gadolinium-enhanced MRI can detect early reversible ischemia of the femoral head epiphysis caused by hip hyper-abduction in piglets. Between 3 and 6 h consistent hyper-abduction, gadolinium-enhanced MRI was performed in 20 femoral heads of 10 piglets. After completion of MRI scan, the piglets were allowed to ambulate freely for 1 or 7 days and re-imaged. The enhanced-MRI results of epiphyseal and physeal cartilage and the secondary center of ossification were observed. MRI appearances and histological findings were compared. On Gadolinium-enhanced MRI, decreased or absent enhancement was seen in 14 cartilaginous epiphyses of all 20 femoral heads. Reperfusion was completed in 10 of 14 femoral heads after one day of ambulation and in the rest 4 after 7 days of ambulation. Gadolinium-enhanced MRI can identify early ischemia and its reversal of the capital femoral epiphysis induced by hip hyper-abduction.

We isolated and identified the symbiotic and adnascent microorganisms from an unidentified sponge collected from 10-meter-deep seawater of the Paracel Islands in China. A total of 16 strains were obtained and identified. Through bacteriostatic activity assay, one of the strains, Dermacoccus sp. X4, was found to effectively inhibit the growth of Staphylococcus aureus. Subsequently, its secondary metabolites were purified by silica gel partition, octadecylsilane (ODS) reverse phase, Sephadex™LH-20 size exclusion, and C18 reverse phase chromatography. Using liquid chromatography, mass spectrometry, and nuclear magnetic resonance, three of the purified compounds were structurally characterized to be one 3-(4-hydroxybenzyl) hexahydropyrrolo [1,2-a]pyrazine-1,4-dione and two indole acid glycerides. This is the first report about indole acid glyceride isolated from microbial secondary metabolites, enriching marine drug candidate resources.
Actinomycetales ; chemistry ; Animals ; China ; Chromatography, Liquid ; Indoles ; isolation & purification ; pharmacology ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Porifera ; microbiology ; Seawater ; Secondary Metabolism ; Staphylococcus aureus ; drug effects

Objective: To establish the culture technique for culturing γδ T cells in vitro and evaluate the basic characteristics, security and anti-tumor effect of the cultured γδ T cells. Methods: Phytohemagglutinin, zoledronic acid, interleukin-2 and interleukin -7 were used to induce the abundant expansion of peripheral blood mononuclear cells in vitro. Flow cytometry assay, in vitro killing assay and mouse model of human lung cancer were also adopted to assess the characteristics and the anti-tumor effect of cultured γδ T cells. Additionally, the contamination of exogenous agents and the acute toxicity of γδ T cells were determined. Results: After culturing 14-16 days in vitro, the total number of γδ T cells was more than 1.0×10¹⁰. Among these γδ T cells, CD3⁺ γδ TCR⁺ cells accounted for more than 90%. None of contaminations of bacteria, fungi, mycoplasma and virus were observed. At effect target ratio (E/T ratio) of 50/1, killing efficiency of γδ T cells cultured in vitro to SK-MES-1, Ho8910, A549 and K562 reached more than 65%. In vivo experiments showed that the tumor volume of γδ T-treated mice was (828.99±61.05) mm^3, significantly lower than (1 723.51±84.30) mm³ of the control mice (P<0.05). Meanwhile, no acute toxicity effect was observed in γδ T cells treated mice. Conclusion The number, purity and activity of γδT cells cultured in our institute can reach the requirement of clinical application, and the γδT cells also display strong cytotoxic activity against tumor cells such as lung cancer, ovarian cancer and leukemia.

Aim To investigate the effect of quercetin on the osteogenic ability of human dental pulp mesenchymal stromal cells (hDPSCs) in vitro and in vivo. Methods hDPSCs were obtained from the pulp tissues of premolar, and the characteristic surface antigens were identified by flow cytometry. Cell Counting Kit-8 (CCK-8) assay was used to test the cytotoxicity of quercetin. Alkaline phosphatase (A L P) and alizarin red staining were used to detect the osteogenic ability of cells in vitro. The expression of osteogenic genes was detected by qPCR. Four round calvarial bone defects with a diameter of 8 mm were created in 10 male New Zealand rabbits, and they were differentiated and randomized into four groups. Group A, hDPSCs cultured on Bio-Oss

Objective: To construct a mouse derived pcDNA3. 1 ⁃3 × Flag⁃c⁃NUP85 expression plasmid and observe its effect on expression of inflammation factors in LPS⁃induced RAW264. 7 cells , as well as on the proliferation and apoptosis of RAW264. 7 cells. Methods: The NUP85 gene was amplified by PCR to construct pcDNA3. 1 ⁃3 × Flag-c⁃NUP85 eukaryotic expression plasmid. The pcDNA3. 1 ⁃3 × Flag⁃c vector was divided with enzymes. The purified PCR product was ligated with the vector, and the ligated product was transformed into bacterial competent cells. After identification by enzyme digestion , sequencing and analysis were performed. Then , it was transfected into RAW264. 7 cells , and the blank plasmid without NUP85 gene was set as the control group. The effect on cell proliferation and apoptosis were detected by CCK⁃8 assay and flow cytometry , and the expression of inflammatory cytokines such as tumor necrosis factor⁃α (TNF⁃α ) and interleukin⁃6 (IL⁃6) in LPS⁃induced RAW264. 7 cells was detected by Western blot and ELISA. Results: Enzyme digestion identification and Western blot results showed that pcDNA3. 1 ⁃3 × Flag⁃c⁃NUP85 eukaryotic expression plasmid was successfully constructed and expressed. The results of CCK⁃8 assay showed that the cell survival rate of NUP85 overexpression group was significantly lower than that of control group after 24 h[(0. 55 ± 0. 03) vs (0. 67 ± 0. 05) , F = 30. 98 , P < 0. 05 ] . The results of flow cytometry showed that the cell apoptosis rate of NUP85 overexpression group was higher than that of control group[( 15. 78 ±1. 05)% vs ( 13. 40 ± 0. 47)% , F = 75. 38 , P < 0. 05] . The results of Western blot and ELISA showed that after transfection of pcDNA3. 1 ⁃3 × Flag⁃c⁃NUP85 , the expression of TNF⁃α and IL⁃6 in RAW264. 7 cells were higher than those in the control group ,with statistical significance (P < 0. 05) . Conclusion NUP85 can inhibit the proliferation and promote apoptosis in LPS⁃stimulated RAW264. 7 cells , and NUP85 can promote the expression of inflammatory cytokines IL⁃6 and TNF⁃α in LPS⁃stimulated RAW264. 7 cells.