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Obｊective To investigate the protective mechanism of tricholoma matsutake polysaccharides(TMP) against 1-methy-4-pehnyl-pyridine ion (MPP
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Objective: To observe the effect of moxibustion on the colonic mucosal barrier of rats with ulcerative colitis (UC) induced by dextran sulfate sodium (DSS). Methods: Forty male Sprague-Dawley rats were randomly divided into a normal group and a modeling group, with 20 rats in each group. Rats in the modeling group were subjected to preparing experimental UC models by drinking 4％ DSS for seven consecutive days. Two modeled rats and two normal rats were randomly selected for model identification. After the success of UC model was confirmed, the remaining 18 modeled rats were randomly divided into three groups, a model group, a model + herbal cake-partitioned moxibustion group, and a model + mild moxibustion group, with six rats in each group; the remaining normal rats were randomly divided into three groups, a normal group, a normal + herbal cake-partitioned moxibustion group, and a normal + mild moxibustion group, with six rats in each group. After 7 d of intervention with the herbal cake-partitioned moxibustion or the mild moxibustion, hematoxylin-eosin (HE) staining technique was used to observe the pathological changes of colon tissue under a light microscope; Western blotting and/or immunohistochemical techniques were used to detect the protein expression levels of Occludin, Claudin, junction adhesion molecular 1 (JAM1), mucin 2 (MUC2), and transforming growth factor beta1 (TGF-β1) in rat colon tissue. Results: Compared with the normal group, the colon tissue was severely damaged, the pathological score was significantly increased, and the protein expression levels of Occludin, Claudin, JAM1, MUC2, and TGF-β1 were significantly decreased in the model group (P<0.01); while there were no significant differences in the colonic histopathological score, protein expression levels of Occludin, Claudin, JAM1, MUC2, and TGF-β1 in the normal + herbal cake-partitioned moxibustion group and the normal + mild moxibustion group (P>0.05). Compared with the model group, the model + herbal cake-partitioned moxibustion group and the model + mild moxibustion group showed repaired colon tissue, ulcer healing, significantly reduced pathological score, and significantly increased protein expression levels of JAM1, MUC2, and TGF-β1 (P<0.05); the Occludin protein expression level in the colon tissue of the model + mild moxibustion group was increased (P<0.01). Conclusion: Neither herbal cake-partitioned moxibustion nor mild moxibustion influences the colonic histopathology and intestinal mucosal barrier-related protein expression in the normal rats; both herbal cake-partitioned moxibustion and mild moxibustion can up-regulate the protein expression levels of JAM1, MUC2, and TGF-β1 in the colon tissue of UC rats. Mild moxibustion can up-regulate Occludin protein expression. This may be a mechanism of moxibustion in reducing colonic mucosa inflammation in UC.
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Objective:To investigate the effect of internet nursing model on female patients who are undergoing in vitro fertilization-embryo transfer, improve the quality of nursing service in reproductive center.Methods:A total of 201 female patients undergoing in vitro fertilization-embryo transfer from April 2018 to December 2018 were divided into the intervention group(102 cases) and the control group(99 cases) by random digits table method. The patients in intervention group were treated with nursing care based on the internet platform, and the control group was given routine care. The duration of intervention was from the patients determined treatment plan to the time pregnancy test 14 days after embryo transfer. The Self-rating Anxiety Scale(SAS) and Self-rating Depression Scale(SDS), Fertility Quality of Life(FertiQoL), and Connor-Davidson Resilience Scale(CD-RISC) were used to evaluate the differences in anxiety and depression, quality of life, and psychological resilience before and after intervention. The outcome of in vitro fertilization-embryo transfer was observed in two groups.Results:After intervention, the score of SAS, SDS were 26.03±6.43, 28.79±5.26 in the intervention group, and 40.76±6.81, 36.13±6.27 in the control group, the difference were significant( t values were -15.77, -9.00, all P<0.01). After intervention,the dimensional scores of mental and physical, social relationship, tolerance to treatment in the FertiQoL were 68.21±18.34, 75.56±19.58, 69.14±18.91 in the intervention group, and 58.11±17.42, 67.24±19.27, 58.77±19.86 in the control group, the difference were significant( t values were 4.00, 3.04, 3.79, all P<0.01). After intervention,the dimensional scores of tough, optimistic in CD-RISC were 37.03±10.51, 14.02±3.25 in the intervention group, and 26.73±9.81, 10.13±3.41 in the control group, the difference were significant( t values were 7.17, 8.28, all P<0.01). The average number of high-quality embryos and clinical pregnancy rate were 1.79(0.59, 4.51), 49.02%(50/102) in the intervention group, and 1.22(0.36, 2.86) , 33.33% (33/99) in the control group, the difference were significant ( t value was 2.47, χ2 value was 5.10, all P<0.05). Conclusions:The new nursing model based on internet has positive significance for the treatment of female patients undergoing in vitro fertilization-embryo transfer, which can improve patients' anxiety and depression, improve their quality of life and enhance their psychological resilience.
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The study aims to investigate the effects of cardiac fibroblast (CF) paracrine factors on murine embryonic stem cells (ESCs). Conditioned mediums from either neonatal cardiac fibroblasts (ConM-NCF) or adult cardiac fibroblasts (ConM-ACF) were diluted by 1:50 and 1:5, respectively, to investigate whether these conditioned mediums impact murine ESCs distinctly with RT-real time PCR techniques, cell proliferation essay, ELISA and by counting percentage of beating embryoid bodies (EBs) during ESCs differentiation. The data showed that the paracrine ability of CFs changed dramatically during development, in which interleukin 6 (IL6) increased with maturation. ConM-NCF 1:50 and ConM-NCF 1:5 had opposite effects on the pluripotent markers, although they both reduced mouse ESC proliferation. ConM-ACF 1:50 promoted ESCs pluripotent markers and proliferation, while ConM-ACF 1:5 exerted negative effects. All CF-derived conditioned mediums inhibited cardiac differentiation, but with distinguishable features: ConM-NCF 1:50 slightly decreased the early cardiac differentiation without altering the maturation tendency or cardiac specific markers in EBs at differentiation of day 17; ConM-ACF 1:50 had more significant inhibitory effects on early cardiac differentiation than ConM-NCF 1:50 and impeded cardiac maturation with upregulation of cardiac specific markers. In addition, IL6 neutralization antibody attenuated positive effect of ConM-ACF 1:50 on ESCs proliferation, but had no effects on ConM-NCF 1:50. Long-term IL6 neutralization reduced the percentage of beating EBs at early developmental stage, but did not alter the late cardiac differentiation. Taken together, both the quality and quantity of factors and cytokines secreted by CFs are critical for the ESC fate. IL6 could be a favorable cytokine for ESC pluripotency and the early cardiac differentiation.
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Animals , Mice , Embryonic Stem Cells , Fibroblasts , Heart , Mouse Embryonic Stem Cells , Paracrine CommunicationABSTRACT
The aim of this study is to investigate the effect of nicotinamide (NIC) on the solubility/dissolution of a poorly soluble drug ibuprofen (IBU), and to explore the mechanism of the formed soluble complex by complexation model, fluorescence quenching and Raman spectroscope. The results showed that NIC could significantly improve the solubility of IBU, and exhibited an Ap type complexation profile. The calculated complexation constants of K1︰1 and K1︰2 were 0.24 and 4.00, respectively. In the solution, the fluorescent intensity of IBU gradually decreased with the increase of NIC, exhibiting the typical fluorescence-quenching phenomenon. The Raman spectrum showed stretching vibrations, bending vibrations, and rocking vibrations ascribed to benzene ring of IBU and pyridine ring of NIC disappeared or significantly shifted, suggested that the soluble complex was formed by dipole-dipole interaction force between the benzene group on IBU and pyridine group on NIC, resulting in the aqueous solubility enhancement of IBU. In comparison to IBU alone, the physical mixture of IBU and NIC showed significantly higher dissolution rate (1.6-fold) and extent.
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Nonsense-mediated mRNA decay (NMD) is originally identified as a widespread mRNA surveillance machinery in degrading 'aberrant' mRNA species with premature termination codons (PTCs) rapidly, which protects the cells from the accumulation of truncated proteins. Recent studies show that NMD can also regulate the degradation of normal gene transcripts, which execute important cellular and physiological functions. Therefore, NMD is considered as a highly conserved post-transcriptional regulatory mechanism in eukaryotes. NMD modulates 3% to 20% of the transcriptome from yeast to human directly or indirectly, which is essential for various physiological processes, such as cell homeostasis, stress response, proliferation, and differentiation. NMD can regulate the level of transcripts that involves in development, and single knockout of most NMD factors has an embryonic lethal effect. NMD plays an important role in the self-renewal, differentiation of embryonic stem cells and is critical during embryonic development. In this review, we summarized the latest advances in the roles and mechanisms of NMD in embryonic development, in order to provide new ideas for the research on embryonic development and the treatment of embryonic development related diseases.
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Humans , Codon, Nonsense , Embryonic Development , Nonsense Mediated mRNA Decay , RNA, Messenger , TranscriptomeABSTRACT
Objective To investigate and evaluate the effects and safety of the operationabsorption of irrigation fluid occurs during 1470 nm diode laser vaporization of the prostate by ethano. Methods We retrospectively analyzed the clinical data about 32 cases of BPH treated by 1470 nm diode laser vaporization from May 2016 to December 2016 intraoperative use of isotonic saline contains 1.0％ ethanol as perfusion fluid, monitor the patients' breath ethanol concentration, calculation of perfusion fluid absorption. Results All the operations were successfully completed, the average operation time was (57.6 ± 32.9) min, the average perfusion fluid volume was (21.5 ± 9.9) L, only 5 cases detected perfusion fluid absorption, the average absorptive amount was (156.8 ± 111.7) ml. During operation, all the patients with stable breathing, circulation, no alcohol poisoning symptoms, no bleeding, capsular perforation, cardiorespiratory failure occurred. All the 32 cases patients were followed up for 3 months, which revealed a significant improvement in dysuresia, no severe complications such as urethrostenosis, urinary incontinence, secondary hemorrhage were noted. Conclusion Fluid absorption little occurs during 1470 nm diode laser vaporization, it is safe and effective in treatment of benign prostatic hyperplasia.
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Objective To find the association between follicular output rate and clinical outcome in patients with polycystic ovarian syndrome (PCOS).Methods A total of 285 female patients with PCOS who were undergoing in vitro fertilization-embryo transfer (IVF-ET) for the first time were included in the study and were divided into three groups according to follicular output rate (FORT) values.The general information,the parameters of the controlled ovarian hyperstimulation,and clinical outcomes were compared.Results The concentration of serum E2 on the day of HCG,preovulatory follicle count (PFC),the number of retrieved oocytes,good-quality embryo rates,clinical pregnancy rate decreased sequently in groups with high,mediate and low FORT value.The incidence of moderate and severe ovarian hyper-stimulation syndrome (OHSS) in the group with mediate FORT value was significantly lower than that of groups with high and low FORT value.The incidence of severe OHSS in 3 groups with high,mediate and low FORT value was 20.6％,9％ and 17.8％ respectively and the difference was statistically significant (P =0.037).Conclusions The value of FORT may predict the clinical outcome of IVF/ ICSI-ET in patients with PCOS.Effort should be made to prevent OHSS in PCOS patients with high or low FORT value in their controlled ovarian hyperstimulation cycles.
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Ursolic and oleanolic acids antagonize ox-LDL-,C-reactive protein-,non-enzymatic glycation end products-,hyper-glucose-,H2O2-,lipopolysaccharide-induced endothelial cell injury,and their vascular pharmacologic effects are in many ways.Ursolic and oleanolic acids can inhibit the proliferation of vascular smooth muscle cells,and antagonize serum-,ox-LDL-,hyper-glucose-,leptin-induced vascular smooth muscle cell proliferation,thus produce vascular protection and improve vascular function;Ursolic and oleanolic acids can relieve diabetic vascular complication in diabetic mellitus rats,and vascular stenosis induced by carotid balloon catheter injury,and suppress the proliferation of vascular adventitial fibroblasts and prevent vascular stenosis induced by various experimental atherosclerosis models;they can also inhibit the proliferation of vascular endothelial cells,and have a dual action to antagonize inhibiting or promoting proliferation induced by various injurious factions.Thus they have a dual role of regulation in angiogenesis,and suppress angiogenesis of cornea,diabetic retina,and tumor tissues,have effects of vascular relaxation and resistance decrease,and have hypotensive effects in normal and various hypertensive animals.
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Oleanolic acid has a widespread pharmacologic effect,including anti-inflammation,antitumor,anfidiabetes,anti-atherosclerosis,hepatoprotection,anti-osteoporosis,etc.Bioavailability of oleanolic acid by oral administration is low,and is absorbed by intestine through passive transport.Oleanolic acid dispersion preparation can increase its solubility and bioavailability,and concentrate on liver to help treat for hepatic disease.The pharmacokinetic parameters,process in body of oleanolic acid,and the pharmacokinetic feature of oleanolic acid dispersion and its prodrug are reviewed,and its research advance on pharmacokinetics is summarized,which provides a reference for rational utilization of oleanolic acid.
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Some animal model experiments have found that salidroside has protective effect for injuries in various neurons and brain tissue induced by various causes,so it is expected to be used in the prevention and treatment of peripheral nerve injury,and cerebral ischemic and neurodegenerative diseases,such as cerebral infarction,Alzheimer's disease,Parkinson's disease,epilepsy,depression,Tourette syndrome,amyotrophic lateral sclerosis,and diabetic encephalopathy.Its mechanisms of neuroprotection are multiple:(1) Salidroside protects neurons and stem cells from apoptotic injury by antioxidation,blocking NOX2/ROS/MAPKs and REDD 1/mTOR/p70S6K signaling pathways,and promoting AMPK/SIRT1,RhoA-MAPK and PI3K/Akt signaling pathways against various injurious factors-induced oxidative stress,inhibiting expression of inflammatory cytokines,preventing intracellular Ca2+ overload and caspase-3 activation;(2) Salidroside obstructs endogenous amyloid-β production by inhibition of expression of β-site amyloid precursor protein cleaving enzyme 1 and β-secretase activity.(3) Salidroside accelerates repair,regeneration and functional recovery of nerve by block Notch signaling pathway,and promoting BMP signaling pathway,super-regulating expression of neurotrophic factors,inducing neural stem cells and mesenchymal stem cells to differentiate into neural cells,and improving proliferation and function of Schwann cells.
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Some animal model experiments have found that salidroside has protective effect for injuries in various neurons and brain tissue induced by various causes,so it is expected to be used in the prevention and treatment of peripheral nerve injury,and cerebral ischemic and neurodegenerative diseases,such as cerebral infarction,Alzheimer's disease,Parkinson's disease,epilepsy,depression,Tourette syndrome,amyotrophic lateral sclerosis,and diabetic encephalopathy.Its mechanisms of neuroprotection are multiple:(1) Salidroside protects neurons and stem cells from apoptotic injury by antioxidation,blocking NOX2/ROS/MAPKs and REDD 1/mTOR/p70S6K signaling pathways,and promoting AMPK/SIRT1,RhoA-MAPK and PI3K/Akt signaling pathways against various injurious factors-induced oxidative stress,inhibiting expression of inflammatory cytokines,preventing intracellular Ca2+ overload and caspase-3 activation;(2) Salidroside obstructs endogenous amyloid-β production by inhibition of expression of β-site amyloid precursor protein cleaving enzyme 1 and β-secretase activity.(3) Salidroside accelerates repair,regeneration and functional recovery of nerve by block Notch signaling pathway,and promoting BMP signaling pathway,super-regulating expression of neurotrophic factors,inducing neural stem cells and mesenchymal stem cells to differentiate into neural cells,and improving proliferation and function of Schwann cells.
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<p><b>OBJECTIVE</b>To explore the relationship between vitamin D receptor (VDR) gene pol-ymorphisms at Fok I site and the risk of preterm birth for potential intervention of of preterm birth or threatened premature delivery.</p><p><b>METHODS</b>Fifty-seven women with preterm birth and 84 with full-term birth were included in this analysis. Polymerase chain reaction-restriction frag-ment length polymorphism (PCR-RFLP) was performed to identify VDR gene Fok I geno-types.</p><p><b>RESULTS</b>No significant difference was found in age, D-dimer (DDI), fibrinogen (Fg), serum calcium (Ca), leukocyte count or glycosylated hemoglobin (HbA1c) level between the women in the preterm and full-term birth groups (P>0.05). The two groups differed signifi-cantly in the distribution of VDR gene Fok I site genotypes and allele frequency of F/F (P<0.05).The frequency of FF genotype was significantly higher in the preterm group than in the full-term group. Compared with Ff and ff genotypes, FF genotype was associated with an in-creased risk of preterm delivery (χ=9.701, P=9.701, OR=3.320, 95% CI [1.560, 7.066]). In the preterm group, the maternal age, DDI, Fg, serum Ca, leukocyte count or HbA1c did not differ significantly between the genotypes (P>0.05).</p><p><b>CONCLUSION</b>VDR gene Fok I site geno-types are related with preterm birth, and the FF genotype may serve as a potential risk factor for preterm birth.</p>
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Objective To investigate the relationship between OHSS and FSHR gene polymorphism. Methods Two hundred and two women were enrolled in this study. The FSHR gene polymorphisms at position 307 and 680 were detected. Results The distribution of the allele frequency and genotype frequency of the position 680 in FSHR gene were significantly different between women with OHSS or not. No significant differences of the position 307 in FSHR gene were observed. Conclusion Themutation of Asn680Ser in FSHR might be closely related with OHSS.
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<p><b>OBJECTIVE</b>To search for the bone mesenchymal stem cell (MSC) subgroup which might be more effective on repairing myocardial damage.</p><p><b>METHODS</b>In this experiment, four MSC subgroups were defined based on the surface differentiation antigen detection of mouse bone mesenchymal stem cells (mBMSCs): SCA-1(+)/CD45(+)/CD31(+), SCA-1(+)/CD45(+)/CD31(-), SCA-1(+)/CD45(-)/CD31(-) and SCA-1(+)/CD45(-)/CD31(+). These subgroup cells and unselected mBMSCs were injected into infarcted mouse via tail vein. Echocardiographic heart function measurement and in vivo DiR-labeled stem cells imaging were performed at 48 h after injection. In situ C-kit (a flag antigen of cardiac stem cells) and cardiac-specific differentiation antigen immunohistochemistry detection was made in the infarcted myocardium.</p><p><b>RESULTS</b>The capacity of the SCA-1(+)/CD45(+)/CD31(+) cells on improving heart function was significantly higher than other cell groups (all P < 0.05). In vivo imaging showed that the mean fluorescence intensity of the SCA-1(+)/CD45(+)/CD31(+) cells was also higher than other cell groups (all P < 0.05). Number of cardiac stem cells in the infracted myocardium was significantly increased after the injection of all subgroup cells and unsorted mBMSCs cells for 48 h compared untreated infracted myocardium. The capacity of mobilizing cardiac stem cells is as follows: SCA-1(+)/CD45(+)/CD31(+) >SCA-1(+)/CD45(-)/CD31(+) >SCA-1(+)/CD45(-)/CD31(-) >SCA-1(+)/CD45(+)/CD31(-).</p><p><b>CONCLUSION</b>The SCA-1(+)/CD45(+)/CD31(+) subgroups of mBMSCs exhibites the highest capacity to improve cardiac function after myocardial infarction and to mobilize autologous cardiac stem cells compared with other mBMSCs subgroups and unsorted mBMSCs cells.</p>
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Animals , Mice , Cells, Cultured , Disease Models, Animal , Mesenchymal Stem Cell Transplantation , Mice, Inbred C57BL , Myocardial Infarction , General SurgeryABSTRACT
Objective To compare the efficacy and tolerability between nonpenetrating filtering surgery with (NPFS-MMC) and without (NPFS-noMMC) intraoperative mitomycin C application for treatment open angle glaucoma. Methods Pertinent studies were selected through extensive searches of the Cochrane Library, PubMed, Embase, and Chinese Biomedicine Database. Eight controlled clinical trials meeting the pre-defined criteria were systematically reviewed by meta-analysis. The main outcome measures were percentage intraocular pressure reduction (IOPR%), complete remission rate, total remission rate, and complications. The pooled estimates were carried out using RevMan version 5.0 software. Results The weighted mean differences of the IOPR% between the NPFS-MMC group and NPFS-noMMC group were 5.24% (95% confidence intervals\[95%CI\], -3.24 to 13.72) after 6 months, 8.31% (95%CI,4.33 to 12.30) after 12 months(P<0.05), 9.56% (95%CI, 4.88 to 14.24) after 24 months(P<0.05), and 14.45% (95%CI, 9.02 to 19.88) after 36 months(P<0.05). NPFS-MMC was associated with significant greater complete remission rates compared with NPFS-noMMC, with the pooled risk difference being 1.16 (95%CI, 1.05 to 1.27) after 6 months(P<0.05), 1.20 (95%CI, 1.05 to 1.38) after 12 months (P<0.05), 1.30 (95%CI, 1.05 to 1.61) after 24 months (P<0.05), and 1.36 (95%CI, 1.06 to 1.73) after 36 months (P<0.05). Intraoperative mitomycin C was not associated with any drug-induced complications. Conclusion The use of intraoperative mitomycin C is safe and can improve the effect of nonpenetrating filtering surgery in patients with open ganle glaucoma.
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<p><b>OBJECTIVE</b>To develop a new method to detect anti-Hantavirus IgG antibodies (HV IgG) based on quantum dots (QDs) and indirect immune technique.</p><p><b>METHODS</b>The carbodiimide crosslinking method was used to couple protein G and goat antihuman IgG on the surface of water-solubility QDs. The coverglass covered HV antigen was used as carrier, and QDs-PG-IgG conjugates was used as labeled second antibody to detect the HV-IgG in the serum samples. The detecting conditions were optimized.</p><p><b>RESULTS</b>The optimum reaction time, pH and goat antihuman IgG concentration for conjugating the QDs with goat antihuman IgG were 6.0, 2h, and 20 microg/ml, respectively. The optimum working dilution of QDs-PG-IgG conjugates was 1: 200. The detection limit of the serum samples was about 1: 1280 dilution.</p><p><b>CONCLUSION</b>The method established in this study has been demonstrated to be a specific, sensitive, rapid test for detecting HV antibodies, laying the foundation of single molecule detection. The anti-fluorescence quenching ability of this method was significant improved.</p>
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Humans , Antibodies, Viral , Blood , Fluorescence , Hantavirus Infections , Diagnosis , Immunoassay , Methods , Immunoglobulin G , Blood , Quantum DotsABSTRACT
<p><b>OBJECTIVE</b>To sum up the experience of performing ascending aorta replacement combined triple-branched stent graft implantation for acute Stanford type A aortic dissection.</p><p><b>METHODS</b>From January 2010 to December 2010, 14 patients with acute Stanford type A aortic dissection underwent the procedure of performing ascending aorta replacement combined triple-branched stent graft implantation. Right axillary artery cannulation was used for cardiopulmonary bypass and selected cerebral perfusion. When the body temperature drops below 18°C, the ascending aorta was transected near the base of the innominate artery. From the incision, the triple-branched stent graft was implanted into the true lumen of the arch, descending aorta and the aorta bifurcation vessel. The transected stump of the ascending aorta was anastomosis to the proximal of the branched blood vessel prosthesis.</p><p><b>RESULTS</b>Cardiopulmonary bypass time was (186 ± 38) min, cross clamp time was (101 ± 27) min, and average selective cerebral perfusion and lower body arrest time was (39 ± 11) min. The in-hospital mortality was zero. One patient of transient postoperative neurologic dysfunction, one of acute renal failure, one of transient limbs disturbance, one of secondary thoracotomy operation, one of gastrointestinal hemorrhage and one of postoperative chylothorax were observed. CT angiography rechecked showed the position of the vascular stent were satisfactory and the blood flow of arterial branches stents were lucid. The false lumen of the aortic arch and descending aorta closed with thrombus or shrinked.</p><p><b>CONCLUSIONS</b>The patients required aortic arch to be reconstructed which had no main tearing of intima in the arch may be best candidates for this technique. Open triple-branched stent graft placement combined ascending aorta replacement is an effective means for aortic arch reconstruction in acute Stanford type A aortic dissection.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Aortic Dissection , General Surgery , Aorta, Thoracic , General Surgery , Blood Vessel Prosthesis , Blood Vessel Prosthesis Implantation , Methods , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To investigate serum IL-18 levels in mice with collagen-induced arthritis treated by recombinant adenoviral vector containing mIL-18BP and mIL-4 fusion gene (AdmIL-18BP/mIL-4).</p><p><b>METHODS</b>Arthritis was induced by injection of collagen in male DBA-1/BOM mice. Mice with collagen-induced arthritis (CIA) were intra-articularly injected with 10(7)pfu/6μL of AdmIL-18BP/mIL-4; and in mice of control groups AdLacZ or PBS were used. The animals were sacrificed at week 1, 2 and 4 after treatment. Serum IL-18 levels were determined by ELISA at the different time points.</p><p><b>RESULT</b>The mean serum levels of IL-18 at weeks 1, 2, and 4 after injection of AdmIL-18BP/mIL-4 were (36.5±5.4)ng/L, (32.5 ± 3.2) ng/L and (28.7 ±2.9)ng/L, respectively, which were significantly lower than those at the same time point of AdLacZ group [(66.2 ±5.1)ng/L, (69.2 ±4.2)ng/L and (77.7 ±3.9)ng/L] and PBS group [(67.3 ±7.1)ng/L, (71.9 ±1.8)ng/L and (78.7±4.1)ng/L] (P<0.01 at all time points). In the therapy group, there were no significant differences in the mean serum concentrations of IL-18 at all time points.</p><p><b>CONCLUSION</b>The serum IL-18 levels in CIA mice are down-regulated by treatment of recombinant adenovirus containing mIL-18BP and mIL-4 fuse gene, which might be a promising therapeutic strategy for rheumatoid arthritis.</p>
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Animals , Male , Mice , Adenoviridae , Genetics , Arthritis, Experimental , Blood , Therapeutics , Gene Fusion , Genetic Therapy , Genetic Vectors , Interleukin-18 , Blood , Genetics , Interleukin-4 , Genetics , Mice, Inbred DBAABSTRACT
<p><b>BACKGROUND</b>Transplantation of adult bone marrow-derived mesenchymal stem cells (MSCs) has been proposed as a strategy for cardiac repair following myocardial damage. However cell transplantation strategies to replace lost myocardium are limited by the inability to deliver large numbers of cells that resist peritransplantation graft cell death. Accordingly, we set out to isolate and expand adult swine bone marrow-derived MSCs, and to engineer these cells to overexpress AKT1 (protein kinase B), to test the hypothesis that AKT1-engineered MSCs are more resistant to apoptosis and can enhance cardiac repair after transplantation into the ischemic swine heart.</p><p><b>METHODS</b>The CDS (regulation domain of AKT1) AKT1-cDNA fragment was amplified, and MSCs were transfected following synthesis with a pCDH1-AKT1 shuttling plasmid. Western blotting analysis and real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed. Myocardial infarction (MI) models were constructed in Meishan pigs, and cardiac function was evaluated by magnetic resonance imaging (MRI) measurements and echocardiography 4 weeks later. All pigs were assigned to four groups: control (A), DMEM (B), MSC (C), and AKT-transfected (D). MSCs were transfected with the AKT1 gene, and autologous BrdU-labeled stem cells (1 x 10(7)/5 ml) were injected into left anterior descending coronary atery (LAD) of the infarct heart in groups C and D. In group B, DMEM was injected using the same approach. In group A, there was no injection following LAD occlusion. After 4 weeks, cardiac function and regional perfusion measurements were repeated by MRI and echocardiography, and histological characteristics of the hearts were assessed. Connecxin-43 (CX-43), BrdU, and von Willebrand factor (VWF) immunoreactivity was tested using enzyme linked immunosorbent assay (ELISA). Vascular endothelial growth factor (VEGF), transforming growth factor-beta1 (TGF-beta1) were analyzed at the same time.</p><p><b>RESULTS</b>AKT1-cDNA was cloned into pCDH1-MCS1-EF1-copGFP and the sequence was confirmed. AKT mRNA expression was detected at 24 hours after transfection. AKT1 expression in MSCs remained strong after 2 weeks, according to real-time RT-PCR and Western blotting. Prior to cell implantation, end-diastolic left ventricular dimension (EDLVd) increased and stroke volume (SV) decreased in the MI hearts. MRI scans revealed significantly improved cardiac function following implantation, and implanted MSCs prevented thinning and expanding in the infarct region, as well as improved contraction and increased perfusion in all groups compared to control hearts. The left ventricular chamber size was smaller in cell-transplanted hearts than in control hearts. Moreover, group D exhibited significant improvement. The expression of CX-43, BrdU, and VWF could be found in the immunohistochemical pathological sections of group C and group D. The level of VEGF reached a high level 1 week after implanting the MSCs, but the level of TGF-beta1 decreased gradually.</p><p><b>CONCLUSIONS</b>The AKT1-expressing lentiviral vector resulted in stable over-expression of AKT1 in MSCs. MSC engraftment in host myocardium improved cardiac function by attenuating contractile dysfunction and pathological thinning of the infracted left ventricular wall, which likely resulted from myocardial regeneration and angiogenesis.</p>