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Objective To explore the effect of exercise on the celluar immune function of mice induced by PM2.5. Methods Twenty-four male C57BL/6 mice were randomly assigned into four groups, filtered air, concentrated PM2.5, exercise and filtered air, and exercise and concentrated PM2.5, with six mice in each group.The mice in the exercise group ran on the treadmill for 1 h every day, and then the mice in the four groups were respectively exposed to concentrated PM2.5 or filtered air after 1 h of rest.After twenty-four weeks, flow cytometric assay was used to detect the quantity of CD3+CD4+CD8a+ T cells, CD18+ T cells and CD154+ T cells in spleen in mice of the four groups. Results The median concentration of PM2.5 in the exposure chamber, filtered chamber and ambient air were 49.24, 12.18, 32.25 μg/m3, respectively.PM2.5 exposure led to increase in CD3+CD4+CD8a+ T cells (P < 0.05) and decrease in CD154+ T cells (P < 0.05), whereas running was beneficial for decreasing CD3+CD4+CD8a+ T cells (P < 0.05) and increasing CD154+ T cells (P < 0.05). Conclusions PM2.5 exposure induces perturbation of the immune system.Regular running proves to be helpful for maintaining the balance of the immune system and improving the body′s resistance to PM2.5.
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Objective To explore the expression of miRLet-7 family members in breast cancer and its correlation with overall survivals (OS), and to find more effective molecular targets for breast cancer prevention and treatment.Methods Kaplan-Meier (KM) plotter online database was used to analyze the correlation among the expression of Let-7 family members (Let-7a, 7b, 7c, 7d, 7e, 7f, 7g, 7i, miR-98, miR-202) correlated with overall survival (OS) and the prognosis and clinical pathological parameters of breast cancer patients, and Hazard ratio (HR), 95％confidence interval (CI), and P value were determined.ResultsThe study showed that the high expression level of Let-7a, Let-7b, Let-7c, Let-7e, Let-7f, Let-7g, miR-98 and the low expression level of miR-202 was associated with better OS for breast cancer patients (P＜0.05).We further assessed the prognostic value of Let-7 in different subtypes and clinical stage.The expression of Let-7a, Let-7b, Let-7f, Let-7g, miR-98, miR-202 was related to clinical stage (P＜0.05).Let-7a, Let-7b, Let-7c, Let-7e, Let-7f, Let-7g, miR-98 and miR-202 was related to lymph node status (P＜0.05).In triple-negative breast cancers (TNBC), with breast cancer subtype, the expression of Let-7b, Let-7c, Let-7g and miR-202 was significantly correlated to overall survival (P＜0.05).Conclusion The Let-7 is significantly correlated with OS in breast cancer patients.The results suggested that members of the Let-7 have different values in predicting the prognosis of breast cancer.Among them, Let-7b, Let-7g and miR-202 are closely related with clinical stage and TNBC, and might promote development of Let-7 as targeted inhibitors for the treatment of breast cancer.
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Objective: To observe the effect of Yiqi Mingmu Pill on the expression of BaxmRNA protein and caspase-3 mRNA in retina of RCS rats. Methods: Twenty-four RCS rats were randomly divided into three groups: blank group, model group, Yiqi Mingmu Pill group. In the blank group, there were 8 RCS (rdy+/+, p+/+) rats, with 4 males and 4 females, and they received intragastric administration of normal saline. The model group consisted of 8 RCS (rdy-/, p-/-) rats, with 4 males and 4 females, and they received intragastric administration of normal saline. Yiqi Mingmu Pills groupconsisted of 8 RCS (rdy-/, p-/-) rats, with 4 males and 4 females, respectively. They received intragastric administrationof suspension of Yiqi Mingmu Pills. After intragastric administration for 30 days, the structure of retina layers wasobserved by HE staining. BaxmRNA and Caspase-3 mRNA in retinal tissues of each group were measured by RT-qPCRand Western Blot. Results: HE staining showed that the retina thickness of rats in Yiqi Mingmu Pill group wassignificantly thicker than that of the model group. The retinal pigment epithelial band was partially visible, and part of theouter nuclear layer photoreceptor sensory ciliary layer was visible. The number of photoreceptor cell nuclei was higherthan that of the model. There were many groups. The outer layer, outer core layer, and the visual cone layer were clearerand thicker than the model group. RT-qPCR showed that the relative expression of Bax mRNA and Caspase-3 mRNA inYiqi Mingmu Pills group was significantly lower than the model group, and the difference was statistically significant (P＜0.01), WB test showed that the relative expression of Bax protein and Caspase-3 protein in Yiqi Mingmu Pill group wassignificantly lower than that in the model group, and the difference was statistically significant (P＜0.01) . Conclusion: YiqiMingmu Pill has protective effect on the ultrastructure of RP retina. Yiqi Mingmu Pills can reduce the apoptosis of retinalphotoreceptor cells by inhibiting the expression of BaxmRNA and Caspase-3 mRNA on the retina and protect the visualcells.
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Objective This study was aimed to analysis the relationship of BK polyomavirus ( BKV) and hemorrhagic cystitis ( HC ) in patients who received hematopoietic stem cell transplantation (HSCT).Methods Data of 80 patients who received HSCT and took regular urine test every week from June 2015 to April 2018 in the Third Xiangya Hospital of Central South University was retrospectively analyzed, they were 35 females and 45 males (aged 20-40 years, median age 30 years), and 31 cases with acute myeloid leukemia ( AML) , 24 cases with acute lymphoblastic leukemia ( ALL) , 15 cases with aplastic anemia ( AA ) , 4 cases with chronic myeloid leukemia ( CML ) , 6 cases with other diseases such as myelodysplastic syndrome ( MDS) in the study population.The positive rate and incidence of HC were analyzed.Patients who infected with the BK virus were divided into HC group and non -HC group according to occurrence of HC.BK viral load were compared in two groups .Urine BK viral load were analyzed after logarithmic transformation.Data that conforms to a normal distribution is expressed as mean ± standard deviation.t test, ANOVA and ROC curve were used to statistical analysis .Skewed data is expressed in median ( interquartile range ) , non-normal distribution parameters were compared by Wilcoxon test .Results Among 80 patients, 43 recipients (53.75％) became urinary BK positive, with 19 patients developed HC (23.75％), all of the 19 HC patients have urinary BK positive , and none of 37 BK-negative patients developed HC;the urine BKV level of the initial time and the peak time in HC group were (7.59 ±2.46) lg copy/ml, (10.56 ±1.71) lg copy/ml, the urine BKV level of the initial time and the peak time in non-HC group were (5.75 ±2.10) lg copy/ml,(7.31 ±2.29) lg copy /ml.The urine BKV level of the initial time and the peak time in HC group was higher than in non-HC group ( t=2.642, P=0.012 and t=5.147, P=0.000 respectively), when analyzing the urine BKV level of the initial time in HC group and non-HC group, the best threshold is 5.23 lg copy/ml(1.68 ×105 copy/ml),with a sensitivity of 84.20％and specificity of 54.17％, when analyzing the urine BKV level of the peak time in HC group and non-HC group, the best threshold is 9.75 lg copy/ml(5.62 ×109 copy/ml),with a sensitivity of 84.20％and specificity of 83.33％, area under curve of each other were 0.728 (95％ CI 0.575-0.881) and 0.875 (95％ CI 0.769-0.981) respectively.Conclusions The BK viral load is closely related with HC in HSCT patients .The cut-off level of 1.68 ×105 copy/ml when analyzing the urine BKV level of the initial time , and the cut-off level of 5.62 × 109 when analyzing the urine BKV level of the peak time , help to forecast or auxiliary diagnose HC .
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TCM experimental teaching has a very important position in university training process, in which the laboratory standardized management and the strengthening of the teaching quality monitoring play important roles. According to the the experimental teaching reforms and requirements enacted in recent years by the Ministry of Education of the People’s Republic of China, Hunan TCM University has conducted a series of standardized management of exploration and practice in the laboratory.
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<p><b>AIM</b>To investigate the dynamic expression of hypoxia-inducible factor 1alpha, PHDs and OS-9 in pulmonary arteries of rats with hypoxia-induced pulmonary hypertension.</p><p><b>METHODS</b>SD rats were randomly divided into 5 groups (n = 8) and exposed to hypoxia for 0, 3, 7, 14 or 21 d, respectively. RT-PCR and in situ hybridization were used to determine the expression of mRNA. Immunohistochemistry and Western blot were used to determine the expression of protein.</p><p><b>RESULTS</b>HIF-1alpha protein was poorly positive in control, markedly up-regulated after 3 d and 7 d of hypoxia (P < 0.05, vs group C), and then declined slightly after 14 d and 21 d of hypoxia. HIF-1alpha mRNA increased dramatically after 14 d of hypoxia (P < 0.05, vs group C). PHD1, PHD2 mRNA and protein was positive in group C. PHD2 mRNA and protein were up-regulated after 3 d of hypoxia (P < 0.05, vs group C), reaching its peak after 14 d of hypoxia while PHD1 protein declined after 14 d of hypoxia (P < 0.05, vs group C) without statistic mRNA changing. PHD3 mRNA and protein were detected at low level in control, markedly up-regulated after 3 d of hypoxia (P < 0.05, vs group C), and then PHD3 mRNA kept at high level while PHD3 protein declined after 14 d of hypoxia (P < 0.05, vs 7 d). OS-9 mRNA was positively in control, markedly decreased after 3 d of hypoxia (P < 0.05, vs group C), reaching its lowest lever after 14 d of hypoxia. Linear correlation analysis showed that OS-9 protein was positively correlated with OS-9 mRNA (r = 0.82, P < 0.01) and HIF-1alpha protein (r = 0.57, P < 0.01).</p><p><b>CONCLUSION</b>HIF-1alpha, PHDs and OS-9 are all involved in the pathogenesis of hypoxic pulmonary hypertension in rats. OS-9 may interact with both HIF-1alpha and PHDs to promote PHD-mediated hydroxylation of HIF-1alpha.</p>
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Animals , Female , Male , Rats , Hypertension, Pulmonary , Metabolism , Hypoxia , Hypoxia-Inducible Factor 1, alpha Subunit , Genetics , Metabolism , Lectins , Genetics , Metabolism , Procollagen-Proline Dioxygenase , Genetics , Metabolism , Pulmonary Artery , Metabolism , RNA, Messenger , Genetics , Metabolism , Random Allocation , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>The underlying mechanisms for cardiac dysfunction in sepsis include the inhibitory effect of endotoxin and inflammatory factors on myocardium and the decrease in cardiac myocardial cells in number. However, whether there is ventricular remodeling resulted from the abnormalities of extracellular collagen metabolism and whether glutamine (Gln) can protect myocardium from LPS-induced damage as in reperfusion are unknown. The aim of the present study was to examine the effects of Gln on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-3 (TIMP-3) and their mRNA in myocardium of rats with sepsis.</p><p><b>METHODS</b>Classical rat model of sepsis was established by intraperitoneal injection of lipopolysaccharide (LPS) (4 mg/kg, from Escherichia coli O(55): B(5), Sigma). from 121 Wistar rats aged 18 days were divided into three groups randomly, 0 h control group (normal saline: 1 ml/kg, n = 11), LPS group (LPS: 4 mg/kg, n = 55) and Gln group (LPS: 4 mg/kg and immediately 13.64% glutamine 1 ml/kg, Fresenus, n = 55). Furthermore, LPS and Gln groups were examined at 2 h, 4 h, 6 h, 24 h and 72 h time points (n = 11). On each time point, rats of LPS and Gln groups as well as control group were anesthetized with 1% chloral hydrate injected intraperitoneally at a dosage of 1 ml/kg. Then, rats were sacrificed, and the hearts were isolated. Eight of them were frozen at minus 80 degrees C to measure the expression of TIMP-3 mRNA by using RT-PCR. The expressions of MMP-3 and TIMP-3 were observed with immunohistochemistry and the expression of MMP-3 mRNA was observed by using in situ hybridization.</p><p><b>RESULTS</b>(1) Compared to 0 h, the mRNA expressions of MMP-3 and TIMP-3 in LPS group significantly increased (P < 0.01) with the peak at 6 - 24 h. While, in Gln group, they were significantly higher than those in controls but significantly lower than those in LPS group with the peak at 24 h (P < 0.01). Even at 72 h, they were still higher than those at 0 h (P < 0.05 and P < 0.01). (2) Compared to 0 h, the expressions of MMP-3 and TIMP-3 in LPS group were significantly lower at any other time point with the lowest at 6 h (P < 0.01). In Gln group, these expressions were also significantly lower than those in controls, but significantly higher than those in LPS group with the lowest being postponed to 24 h (P < 0.01). (3) The ultra structure changed obviously. Z line was unclear and the ridge of mitochondrion disappeared. While, in Gln group, the myocardial injury was slight compared to that in LPS group.</p><p><b>CONCLUSIONS</b>MMP-3 mRNA expression was increased and TIMP-3 mRNA expression was depressed in LPS-induced sepsis. Myocardial extracellular matrix was damaged in sepsis. Glutamine might decrease the effects of LPS on MMP-3 and TIMP-3 expressions and postpone the time of myocardial matrix injury.</p>
Subject(s)
Animals , Female , Male , Rats , Animals, Newborn , Disease Models, Animal , Glutamine , Pharmacology , Immunohistochemistry , In Situ Hybridization , Lipopolysaccharides , Toxicity , Matrix Metalloproteinase 3 , Metabolism , Myocardium , Cell Biology , Metabolism , Myocytes, Cardiac , Metabolism , RNA, Messenger , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Sepsis , Drug Therapy , Genetics , Metabolism , Time Factors , Tissue Inhibitor of Metalloproteinase-3 , MetabolismABSTRACT
@#ObjectiveTo explore the methods of low temperature preservation for alginate-polylysine-alginate (APA) microcapsules.MethodsAPA microcapsules were prepared with static electricity, and underwent hypothermal treatment respectively through methods of program control, gradient by icebox and put in liquid nitrogen directly, finally preserved in liquid nitrogen. The form and permeability of APA microcapsules were checked after rewarming.ResultsThe rates of integrity, crenation and damage were (91.2±1.57)%, (3.1±0.81)% and (5.7± 2.62 )% in the program control group; (85.3±1.42)%, (5.2±0.74)% and (9.5± 3.81 )% in the gradient by icebox group; (14.5±1.57)%, (84.1±3.47)% and (1.4±2.62)% in the directly put in liquid nitrogen group. The membrane permeability of full APA microcapsules after frozen and reworming was not changed obviously.ConclusionThe program control method can preferably preserve APA microcapsules at low temperature and keep them having normal form and permeability.
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<p><b>OBJECTIVE</b>Endotoxemia is a serious syndrome resulting in multi-organ failure. Once it happens, the penetration of small intestine epithelium increases, body liquid losses, then effective circulating blood decreases and serious metabolic acidosis, serious hypotension, systolic failure, and even shock may occur. In this pathological process, endotoxin, tumor necrosis alpha and systolic dysfunction play important roles. Nowadays, many studies have been done to resolve the systolic dysfunction, but too much attention had been paid to the followings: the depressions of myocardium caused by tumor necrosis alpha, other inflammatory factors, endotoxin and metabolic acidosis; the disturbance of blood vessel-nerve regulations; nitric oxide (NO)/inducible nitric oxide synthase (iNOS) over-synthesis and the decreased density of beta-receptors in the myocardium and/or their activities. Little attention has been paid to the relationship between alpha sarcmeric actin (alpha-SA) and systolic dysfunction during endotoxemia. Glutamine (Gln) can be metabolized into glutathione, an eliminator of free radical. It has been used in preventing myocardial damage from reperfusion. This study aimed to observe the dynamic changes of alpha-SA and mRNA expressions in rats with endotoxemia and examine the effects of Gln on them.</p><p><b>METHODS</b>Classical rat model of endotoxemia was established by intraperitoneal injection of LPS (4 mg/kg, Escherichia coli O55:B5, Sigma). 121 Wistar 18-day-rats were divided into three groups randomly, (1) 0 h control group (normal saline: 1 ml/kg, n = 11). (2) LPS group (LPS: 4 mg/kg, n = 55). (3) Gln group (LPS: 4 mg/kg and immediately 13.64%; Gln: 1 ml/kg, Fresenus, n = 55), Furthermore, LPS and Gln groups were divided into 2, 4, 6, 24 and 72 h time points (n = 11). Each time point of LPS and Gln as well as control rats were anaesthetized at each time point with 1% chloral hydrate injected intraperitoneally at the dosage of 1 ml/kg. Then rats were sacrificed at appoint time, and the hearts were isolated. Eight of them were put in 76 degrees C liquid nitrogen and then frozen in minute 80 degrees C icebox in order to measure the expression of alpha-SA mRNA by RT-PCR. Three of them were fixed in 4% formaldehydum polymerisatum for 12 to 16 h, then the expression of alpha-SA was detected by immunohistochemistry.</p><p><b>RESULTS</b>(1) Compared to 0 h, the expressions of alpha-SA and mRNA in LPS group were significantly depressed (P < 0.01). In LPS group, the lowest was at 6 - 24 h, while in Gln group, it was postponed to 24 h. At 72 h, there was no difference in expressions of alpha-SA between Gln and 0 h group (P > 0.05). (2) Comparing at same time point, the expressions of alpha-SA were significant higher in Gln group than those in LPS group, while the expressions of alpha-SA mRNA in Gln group were high at 4-72 h. There was, however, no significant difference at early phase (P > 0.05).</p><p><b>CONCLUSION</b>Alpha-SA and its mRNA expression were depressed in LPS-induced endotoxemia, especially from 6 to 24 h. It could damage the systolic function. alpha-SA decrease in endotoxemia was due to the inhibited synthesis other than the promoted degradation. Glutamine could inhibit the effects of LPS on both alpha-SA and its mRNA expressions.</p>