Study of senescence protein p66Shc on myocardial tissue repair in adult mice / 生理学报

Our previous study has shown that p66Shc plays an important role in the process of myocardial regeneration in newborn mice, and p66Shc deficiency leads to weakened myocardial regeneration in newborn mice. This study aims to explore the role of p66Shc protein in myocardial injury repair after myocardial infarction in adult mice, in order to provide a new target for the treatment of myocardial injury after myocardial infarction. Mouse myocardial infarction models of adult wild-type (WT) and p66Shc knockout (KO) were constructed by anterior descending branch ligation. The survival rate and heart-to-body weight ratio of two models were compared and analyzed. Masson's staining was used to identify scar area of injured myocardial tissue, and myocyte area was determined by wheat germ agglutinin (WGA) staining. TUNEL staining was used to detect the cardiomyocyte apoptosis. The protein expression of brain natriuretic peptide (BNP), a common marker of myocardial hypertrophy, was detected by Western blotting. The results showed that there was no significant difference in survival rate, myocardial scar area, myocyte apoptosis, and heart weight to body weight ratio between the WT and p66ShcKO mice after myocardial infarction surgery. Whereas the protein expression level of BNP in the p66ShcKO mice was significantly down-regulated compared with that in the WT mice. These results suggest that, unlike in neonatal mice, the deletion of p66Shc has no significant effect on myocardial injury repair after myocardial infarction in adult mice.

Vascular Transcriptome Profiling Reveals Aging-Related Genes in Angiotensin Ⅱ-Induced Hypertensive Mouse Aortas / 中国医学科学杂志(英文版)

Objective Angiotensin Ⅱ (Ang Ⅱ)-induced vascular damage is a major risk of hypertension. However, the underlying molecular mechanism of AngⅡ-induced vascular damage is still unclear. In this study, we explored the novel mechanism associated with Ang II-induced hypertension. Methods We treated 8- to 12-week-old C57BL/6J male mice with saline and Ang Ⅱ(0.72 mg/kg·d) for 28 days, respectively. Then the RNA of the media from the collected mice aortas was extracted for transcriptome sequencing. Principal component analysis was applied to show a clear separation of different samples and the distribution of differentially expressed genes was manifested by Volcano plot. Functional annotations including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were performed to reveal the molecular mechanism of Ang Ⅱ-induced hypertension. Finally, the differentially expressed genes were validated by using quantitative real-time PCR. Results The result revealed that a total of 773 genes, including 599 up-regulated genes and 174 down-regulated genes, were differentially expressed in the aorta of Ang Ⅱ-induced hypertension mice model. Functional analysis of differentially expressed genes manifested that various cellular processes may be involved in the Ang Ⅱ-induced hypertension, including some pathways associated with hypertension such as extracellular matrix, inflammation and immune response. Interestingly, we also found that the differentially expressed genes were enriched in vascular aging pathway, and further validated that the expression levels of insulin-like growth factor 1 and adiponectin were significantly increased (P<0.05). Conclusion We identify that vascular aging is involved in Ang Ⅱ-induced hypertension, and insulin-like growth factor 1 and adiponectin may be important candidate genes leading to vascular aging.

IFNα induces cytidine/uridine monophosphate kinase 2 expression in hepatocarcinoma cell line to activate macrophages of mice / 基础医学与临床

Objective To explore the role of cytidine/uridine monophosphate kinase 2( CMPK2) in the immune-mediated antitumor effect of IFNα in hepatocellular carcinoma. Methods RT-qPCR and Western blot were used to analyze the expression of CMPK2 in Huh7 after the treatment of IFNα. The CMPK2 overexpressing Huh7 cells were generated by stably infecting with lentivirus. The ATP level in the cells and the supernatant of CMPK2 overexpress-ing Huh7 cells were measured by CellTiter-Glo ATP fluorescence assay. RT-qPCR was applied to test the expression of inflammatory cytokines in macrophages under the treatment of the supernatant of CMPK2 overexpress-ing Huh7 cells. Results The transcription and protein level of CMPK2 were significantly enhanced after the treat-ment of IFNα for 6 hours ( P＜0.01) . CMPK2 increased the ATP level in the cells and supernatant of Huh7 cells ( P＜0.01) . The supernatant of CMPK2 overexpressing Huh7 cells activated the expression of IL1β, IL6 and CCL5 in macrophages( P＜0.01) . Conclusions IFNα increases the expression of CMPK2 in Huh7 cells to activate the expression of inflammatory cytokines in macrophages.

Impact of rhG-CSF mobilization on TH17/Treg cells in donors' peripheral blood / 中华器官移植杂志

Objective To explore the effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) mobilization on TH17/Treg cells and its impact on suppressor of cytokine signaling-3 (SOCS3) gene expression in CD4+ T cells in donors' peripheral blood.Method Sixteen donors were injected subcutaneously with rhG-CSF 5 μg/kg every day for 5 consecutive days for peripheral blood stem cells mobilization.At the first 0,3,5 day,the mononuclear cclls (MNCs) in peripheral blood or graft and serum specimens were taken.The CD4 + T cells in MNCs were sorted using immuno-magnetic beads.The ratio of TH 17 and Treg cells in MNCs,cytokines concentrations of IL-17A,IL-21,ID23 and TGFβ1 in serum,and SODC3 gene expression in CD4+ T cells were detected by using flow cytometry,ELISA,and reverse transcription real-time quantitative PCR (RT-qPCR),respectively.Results (1)The ratio of Th17 cells (CD3+ CD8 CD17+) and Treg cells (CD4+ CD25+ Foxp3+) in MNCs in peripheral blood and graft at the first 0,3 and 5 days after mobilization was (2.69 ± 0.81) ％,(0.91 ± 0.33) ％,(0.35 ± 0.12) ％,(0.21 ± 0.05) ％,and (0.56 ± 0.24) ％,(0.72 ± 0.22％),(1.59 ± 0.54) ％,(3.38 ± 0.52) ％,respectively,showing a significant declining and increasing trend respectively (P＜0.05); (2)The cytokine concentrations in serum at the first 0,3 and 5 days after mobilization were 7.33 ± 0.89,5.78 ± 1.03 and 3.32 ± 0.84 μg/L for IL-17A; 124.56 ± 15.18,117.12 ± 14.45 and 64.94 ± 11.25 μg/L for IL-21 ; 183.52 ± 59.35,280.49 ± 69.75 and 393.62 ± 57.25μg/L for TGF-β1 (P＜0.01) ; and 45.89 ± 6.95,46.25 ± 7.44 and 47.45 ± 10.75 μg/L for IL-23,respectively.The IL-17A and IL-21 concentrations showed significant declining trend,contrarily TGF-β1 with an increasing trend,while IL-23 concentration had no change.After rhG-CSF mobilization,the SOCS3 gene expression in CD4 + T cells of peripheral blood and graft at the first 0,3,5 days was gradually increased.Conclusion rhG-CSF suppresses Th17 cells and promotes regulatory T cells generation,meanwhile decreases IL-17A and IL-21 and elevates serum TGF-β1 concentrations,and contributes to CD4 + T cells differentiation to Tregs,probably by elevating SOSC3 gene expression in CD4+ T cells.

Up-regulation of Fas ligand expression by sirtuin 1 in both flow-restricted vessels and serum-stimulated vascular smooth muscle cells / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To study the role of sirtuin 1 (SIRT1) in Fas ligand (FasL) expression regulation during vascular lesion formation and to elucidate the potential mechanisms.</p><p><b>METHODS</b>SIRT1 and FasL protein levels were detected by Western blotting in either mouse arteries extract or the whole rat aortic vascular smooth muscle cell (VSMC) lysate. Smooth muscle cell (SMC)-specific human SIRT1 transgenic (Tg) C57BL/6 mice and their littermate wild-type (WT) controls underwent complete carotid artery ligation (ligation groups) or the ligation-excluded operation (sham groups). The carotid arteries were collected 1 day after operation. Reverse transcription-polymerase chain reaction was performed to detect the mRNA levels of SIRT1 and FasL. Luciferase reporter assays were performed to detect the effect of WT-SIRT1, a dominant-negative form of SIRT1 (SIRT1H363Y), and GATA-6 on the promoter activity of FasL. Flow cytometry assay was applied to measure the hypodiploid DNA content of VSMC so as to monitor cellular apoptosis.</p><p><b>RESULTS</b>SIRT1 was expressed in both rat aortic VSMCs and mouse arteries. Forced SIRT1 expression increased FasL expression both in injured mouse carotid arteries 1 day after ligation (P<0.001) and VSMCs treated with serum (P<0.05 at the transcriptional level, P<0.001 at the protein level). No notable apoptosis was observed. Furthermore, transcription factor GATA-6 increased the promoter activity of FasL (P<0.001). The induction of FasL promoter activity by GATA-6 was enhanced by WT-SIRT1 (P<0.001), while SIRT1H363Y significantly relieved the enhancing effect of WT-SIRT1 on GATA-6 (P<0.001).</p><p><b>CONCLUSIONS</b>Overexpression of SIRT1 up-regulates FasL expression in both flow-restricted mouse carotid arteries and serum-stimulated VSMCs. The transcription factor GATA-6 participates in the transcriptional regulation of FasL expression by SIRT1.</p>

Lysine-specific demethylase 1 represses THP-1 monocyte-to-macrophage differentiation / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the role of lysine-specific demethylase 1 (LSD1) in the process of THP-1 monocyte-to-macrophage differentiation.</p><p><b>METHODS</b>Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting were performed to analyze the expression of LSD1 and interleukin-6 (IL-6) in THP-1 monocytes and THP-1-derived macrophages. Chromatin immunoprecipitation (ChIP) assay was applied to detect the occupancy of LSD1 and H3K4 methylation at IL-6 promoter during THP-1 monocyte-to-macrophage differentiation. IL-6 mRNA level and H3K4 methylation at IL-6 promoter were analyzed using qRT-PCR and ChIP assay in LSD1-knockdown THP-1 cells treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) for 0, 4, 8, 12, and 24 hours. Fluorescence activated flow cytometry was performed to reveal the percentage of macrophages differentiated from THP-1 monocytes.</p><p><b>RESULTS</b>The expression of LSD1 reduced during THP-1 monocyte-to-macrophage differentiation (P<0.01). LSD1 occupancy decreased and H3K4 methylation increased at IL-6 promoter during the differentiation. With knockdown of LSD1, H3K4 methylation at IL-6 promoter was found increased after TPA treatment at different times points (all P<0.05, except 24 hours). The percentage of macrophages increased significantly in the THP-1 cells with LSD1 knockdown (P<0.05).</p><p><b>CONCLUSIONS</b>LSD1 is repressed during the monocyte-to-macrophage differentiation of THP-1 cells. Suppression of LSD1-mediated H3K4 demethylation may be required for THP-1 monocyte-to-macrophage differentiation.</p>

Direct secretory expression of active microbial transglutaminase in Pichia pastoris / 生物工程学报

Direct secretory expression of active microbial transglutaminase (MTG) using heterologous hosts is a promising strategy, although its production level still needs to be improved for industrial production. Pichia pastoris is one of the most efficient expression systems developed in recent years. In this study, secretory expression of active MTG was successfully achieved by co-expressing the pro sequence and mature MTG genes in P. pastoris. Furthermore, we optimized the copy number of pro/MTG expression cassettes and the fermentation conditions. MTG production level reached 7.3 U/mL in 1-liter fermentor through high density fermentation, providing the feasiblity for industrial scale preparation of MTG.

Research progression of deacetylase (SIRT1) / 中国医学科学院学报

The silent information regulator protein 2 (Sir2) and its homologues play an important role in the regulation of cellular physiological processes such as survival, apoptosis, and aging. SIRT1, the mammalian Sir 2 homologue, has been shown to deacetylate a wide range of non-histone substrates and histone substrates. It has been constantly reported that SIRT1 may be associated with the occurrence of metabolic syndrome, genomic homeostasis, tumors, and neurodegenerative diseases. Calorie restriction may mitigate many major diseases in rodent models by SIRT1-mediated deacetylase activity and prolong the life expectancies in these animals. Therefore, SIRT1 may be emphasized as a new therapy target for many different diseases.

Observation of the L929 cell membrane after infrasound exposure with atomic force microscope / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the changes of L929 cell membrane with atomic force microscope (AFM) after infrasound exposure and to explore the mechanisms of effect of infrasound on cell membrane.</p><p><b>METHODS</b>After primary culture, the L929 cells were exposed to infrasound with intensity output of 130 dB and frequency of 16 Hz 2 hours each day for 3 days. The subsequent changes in the membrane of the control cells and the cells exposed to the infrasound were determined by nano-scale scanning with AFM.</p><p><b>RESULTS</b>After infrasound exposure, the normal prominence of the membrane became short and the dent became shallow in the 7.5 microm x 7.5 microm and 4.0 microm x 4.0 microm photographs. The prominence appeared as cobblestones. The surface of the membrane became smooth.</p><p><b>CONCLUSION</b>The membrane structure of the L929 cells can be changed by infrasound exposure with intensity of 130 dB and frequency of 16 Hz. The change might be one of the characteristics of effect of infrasound on cell membrane.</p>

Effect of 8 Hz infrasound on expression of 5-HT in rats' hippocampus and temporal cortex / 中国康复理论与实践

@#ObjectiveTo study the effect of 8 Hz infrasound on the expression of 5-hydroxytryptamine (5-HT) in rats' hippocampus and temporal cortex.Methods140 male SD rats were randomly divided into the control group, and experimental groups that exposed to infrasound of 8Hz,90dB,100dB and 130dB for 1,7,14,21,28,35,42 days. Experimental groups were exposed to infrasound for 2 hours each day. The control group was only placed in the infrasonic storehouse but without infrasound. Rats' brains were taken as soon as the exposure finished and strained by immunohistochemistry. The content of 5-HT in hippocampus and temporal cortex was detected under an optical microscope.ResultsInfrasound groups had less expression of 5-HT in hippocampus and temporal cortex than the control group (P<0.01), and the least were at the 28th day for 90 dB and 100 dB groups and the 21st day for 130 dB group. Then the expression of 5-HT had an increase in each group.ConclusionThe deceased expression of 5-HT in rats' hippocampus and temporal cortex could result from infrasound of 8 Hz. Rules of change are related to the parameter of infrasound and the 130 dB 8 Hz infrasound can induce greater changes compared with that of 100 dB and 90 dB.

Effects of infrasound exposure on several enzymes activities of spleen and liver in rats / 中国应用生理学杂志

<p><b>AIM</b>To investigate the changes of several enzymes activities in the spleen and liver of rats after exposure to 8 Hz 130 dB infrasound for different time.</p><p><b>METHODS</b>Thirty-five male SD rats were randomly divided into five groups. Rats of group 1 served as control, rats from group 2 to 5 were exposed to 8 Hz 130 dB infrasound, 2 hours per day, for 1 wk, 2 wk, 3 wk, and 4 wk, respectively. The changes of enzymes activities in spleen and liver of rats were observed.</p><p><b>RESULTS</b>Monoamine oxidase activities in spleen were significantly increased at 1 wk and 2 wk, it was decreased at 3 wk, and increased again at 4 wk (P < 0.05). There were no changes in the liver compared with the control group. Glutathione peroxides activities in spleen were significantly increased at 4 wk (P < 0.05) and it also increased in liver at 1 wk (P < 0.05). Superoxide dismutase activities in spleen were increased significantly from 1 wk to 4 wk, but there were no markedly changes in liver. The level of malondialdehyde in spleen were increased at 3 wk and 4 wk. In the liver, it were increased at 1 wk and 2 wk, and decreased at 3 wk, but it increased again at 4 wk (P < 0.05).</p><p><b>CONCLUSION</b>The results indicated that lipid peroxidation and oxygen free radicals in spleen and liver were increased after infrasound exposure and it might induce the damage in tissue or cells.</p>

Effects of 8 Hz,90 dB/130 dB infrasound on expression of NMDAR1 in hippocampus of rat / 中华物理医学与康复杂志

Objective To study the effect of infrasound on the changes of expression of NMDAR1 in hipp- ocampal cells.Methods Eighty-eight male Sprague-Dawley rats were randomized into eleven groups:control group,90 dB/1 d,7 d,14 d,21 d and 28 d infrasound exposed groups;130 dB/1 d,7 d,14 d,21 d and 28 d infra- sound exposed groups.All the animals in the test groups were put in an infrasound field with 8 Hz,90 dB or 130 dB for 2 hours daily.Immunohistochemistry methods were used to detect the changes of intracellular expression of NMDARI in hippocampal cells.Methods The expression of NMDAR1 in hippocampus after the rats were exposed to infrasound of 8 Hz,90 dB SPL showed a procedure from reducing on the 1st day to rising on the 7th and peaked on the 14th day,then to descending on 21st day and returning to the standard level on the 28th day.Exposure to infra- sound of 8 Hz,130 dB SPL induced opposite effects on the expression of NMDAR1 compared with 90 dB SPL,which showed a process of increasing,descending,reaching to the lowest,then ascending and returning to the normal.The lowest expression of NMDAR1 occurred on the 14th day in every observed hippocampal area.Conclusion 8 Hz, 90 dB/130 dB infrasound induced certain reversible reaction in the expression of NMDAR1 of hippoeampal cells in rats,which may disturb their learning and memory function.

Effects of infrasound on the auditory function of guinea pigs / 中华物理医学与康复杂志

Objective To observe the effects of exposure to infrasound on auditory function of guinea pigs as reflected by the brain stem response (ABR) and distortion product otoaeoustic emission(DPOAE).Methods Seventy-two Guinea pigs were used in this study,of them,12 served as controls and 60 were divided into 2 experi- mental subgroups.ABR and DPOAE were detected after exposure to infrasound stimulation at 16 Hz,90 dB or 16 Hz,130 dB for 2 hours a day for 1 ,7,14,21 and 28 days.Results The threshold of ABR after exposed to infrasound at 16 Hz,90 dB in 1 day and 28 days was higher than the controls (P0.05),but significant difference was observed with 16 Hz 130 dB infrasound exposure on the 21st and,28th days when compared with the controls (P

The effects of infrasound on the morphology of the gastric mucosa / 中华物理医学与康复杂志

Objective To observe the effects of infrasound(IS)of different frequencies and intensities on the pathological morphology of the gastric mucosa(GM)in rats.Methods One hundred and forty male Sprague- Dawley rats were randomly and evenly divided into a control group(group A),an 8 Hz 90 dB group(group B),an 8 Hz 130 dB group(group C)and a 16 Hz 130 dB group(group D).IS with these frequencies and intensities was administered daily for 2 h to all groups except group A,which received sham infrasound.The other 70 rats were ran- domly and evenly divided into a second control group(group E)and an exposure group(group F),in which the rats were continuously stimulated by IS at 8 Hz and 130 dB for 2 h a day for 14 d.The pathological morphology of the GM in each group was observed at 1 d,7 d,14 d,21 d and 28 d after IS exposure.Results①Compared with group A,GM lesion scores were significantly increased in groups B,C and D at 1 d,7 d,14 d,21 d and 28 d(P＜0.01),but not in group B at 1 d.②Compared with group B,the GM lesion scores in group C were obviously in- creased at 1 d,7 d,14 d,21 d and 28 d,while scores were also obviously improved in group C in comparison with those in group D at 14 d,21 d and 28 d(P＜0.01).③The GM lesion scores in group F decreased gradually after IS,but were still higher than those in group E at 1 d,7 d,14 d,21 d and 28 d after IS.④The ultrastructures of the chondrosome and endocytoplasmic reticulum in GM cells were deformed after 8 Hz and 16 Hz IS.Conclusion 8 Hz 90 dB,8 Hz 130 dB and 16 Hz 130 dB IS can all result in GM damage in rats.The injury severity was closely related to the frequency,intensity and duration of the IS.Rats can adapt to IS after several exposures,and the damage tends to recover automatically.

Effect of low-intensity millimeter wave irradiation on the immune adhesion function of erythrocytes in mice with tumor / 中国康复理论与实践

@#ObjectiveTo study the effect of low-intensity millimeter wave(MMW)irradiation on the immune adhesion function of erythrocytes in mice with tumor.Methods60 healthy male mice were randomized into two groups:irradiation group(n=30) and control group(n=30).All mice were injected into abdominal cavity with A's abdominal juice cancer-cell(1×106/ml,0.2ml).MMW(36GHz,0.73-1.46mw/cm2)was used to irradiate the back of mice in irradiation group,once a day for 8 days.Control group received false irradiation using the same method.At 0d,3d,6d after irradiation, the erythrocytes were extracted from vein blood in the mice,and the RBC-C3b receptor rosettes rate(RCR),tumour-RBC rosette rate(TRR),RBC-SOD and RBC-LPO were determined.ResultsAt 3d after radiation,the RBC-C3b and TRR of irradiation group were higher than that of control group (P<0.01) and RBC-LPO was lower (P<0.05),while there was no significant difference on activity of RBC-SOD(P>0.05). there were no significant difference on all index between two groups at 0d and 6d(P>0.05).ConclusionsLow-intensity millimeter wave irradiation can improve the immune adhesion function of erythrocytes in mice with tumor.

Effects of infrasound on ultrastructure of testis cell in mice / 中华男科学杂志

<p><b>OBJECTIVES</b>To investigate the effects of infrasound on ultrastructure of testis in mouse.</p><p><b>METHODS</b>Twelve male BALB/C mice were randomly divided into three groups according to exposed duration on 1, 7 and 14 day. The mice were separately exposed to infrasound environment under 8 Hz/90 dB, 8 Hz/130 dB, 16 Hz/90 dB, 16 Hz/130 dB 2 hours per day. There was another control group which had three mice were separated into module with no infrasound. All the mice were killed on schedule. Then all the sections of testis were observed under electronic microscope. The alterations of structure and the chromatin were observed.</p><p><b>RESULTS</b>Some acute alteration in one day group was found in testis cell, such as cellular denaturation and necrosis, intercellular edema, mitochondria swelling, liposome hyperplasia. When the infrasound was up to 8 Hz/130 dB, the damage showed seriously. In 7 and 14 day group, the acute alteration was gradually decreased. A plenty of abnormal sperm were found. And other alteration was chromatin condense. The effect of variational frequency was important in ultrastructure.</p><p><b>CONCLUSIONS</b>The infrasound markedly effected to testicular cell morphology and secreting function. Infrasound will lead to the alteration of procreation in mouse.</p>