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The paleoenvironment of Jixian carbonate rocks in the Mesoproterozoic Ordos Basin is studied by carbon and oxygen isotope analyses, diagenetic environment analysis, and the restoration of paleosalinity and paleotemperature. The results indicate that the carbonate rocks of the Jixian System have always been in a near-surface environment and have not been deeply buried. The ranges of variation in δ13CPDB and δ18OPDB are relatively narrow, ranging from - 5.75 to 1.41 and - 8.88 to - 4.01, respectively, which is consistent with the stable tidal flat sedimentary environment during the Mesoproterozoic in the study area. The paleosalinity (Z) values range from 111.7 to 127.1, and the paleotemperature (T) values range from 32.7 to 57.33 °C, indicating a relatively warm paleoclimatic environment during the Mesoproterozoic era in the study area. The analysis shows that in a warm paleoclimatic environment, although carbon and oxygen isotopes, Z, and T have certain fluctuations, their ranges are relatively small, reflecting to some extent the stable tectonic environment of the study area during the Mesoproterozoic era. Comprehensive research shows that the Ordos Basin had a warm climate and a stable tectonic environment in the Mesoproterozoic, which may be a good response to the North China Block's position near the equator and continuous thermal subsidence in the Mesoproterozoic.
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Objective:To evaluate the optimization efficacy of ropivacaine mixed with nalbuphine for epidural labor analgesia through comparison with ropivacaine mixed with sufentanil in primiparas.Methods:Four hundred and forty primiparas with a singleton fetus in vertex presentation without abnormal fetus, aged 19-36 yr, at 37-42 weeks of gestation, who were suitable and volunteered to receive epidural labor analgesia, were selected and randomly divided into observation group (NR group, n=220) and conventional group (C group, n=220).Epidural labor analgesia solution contained nalbuphine 40 mg and ropivacaine 120 mg in 150 ml of normal saline in group NR.Epidural labor analgesia solution contained sufentanil 50 μg and ropivacaine 120 mg in 150 ml of normal saline in group C. The analgesia pump was set up to deliver a 5 ml bolus dose with a 20-min lockout interval, programmed intermittent bolus 10-15 ml/h and background infusion at 2 ml/h after an initial dose of 15 ml to maintain VAS score <3.The length of labor, adverse reactions during labor analgesia, neonatal Apgar score, parameters of umbilical artery blood gas analysis and neonatal behavioral neurological assessment score were recorded. Results:Compared with group C, the incidence of urinary retention, pruritus, nausea and vomiting and fever at birth was significantly decreased in group NR ( P<0.05).The duration of the first and second stages of labor, neonatal Apgar score, pH value of umbilical artery blood, lactate concentrations, and neonatal behavioral neurological assessment score were within the normal range in both groups, and there was no significant difference between the two groups ( P>0.05). Conclusions:Compared with conventional medication, nalbuphine mixed with ropivacaine has the same efficacy when used for epidural labor analgesia in primiparas, but it has a certain optimization efficacy in terms of safety.
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Crystalline silica (SiO2) particles have very strong toxicity to the lungs, and silicosis is an excessive pulmonary interstitial remodeling disease that follows persistent SiO2 injury. We showed here that DNA double strand breaks (DSBs) and apoptosis were aggravated during rat silicosis induced by SiO2 exposure. Ac-SDKP attenuates lung parenchymal distortion and collagen deposition, and decreases the expression of γH2AX, p21, and cleaved caspase-3, as well as improves the reduction of pulmonary function caused by silicosis. In vitro, we found an evolution of smooth muscle actin α (α-SMA), collagen type I (Col I) in both A549 and MRC-5 cells in response to transforming growth factor-beta 1 (TGF-ß1)â¯+â¯SiO2. Only A549 cells showed any reduction in the rate of apoptosis induced by the double stimulation, because of the anti-apoptotic effects of TGF-ß1. N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is an anti-fibrotic tetrapeptide. It also has the ability to promote the apoptosis of leukemia cells. However its role in promoting cell apoptosis in silicosis is still unknown. We here found that Ac-SDKP could induce cell apoptosis and inhibit fibrotic response in A549 and MRC-5 cells treated with TGF-ß1â¯+â¯SiO2, and these effects depended on regulation of α-tubulin acetyltransferase 1 (α-TAT1). These findings suggest that Ac-SDKP may have therapeutic value in the treatment of silicotic fibrosis.
Subject(s)
Acetyltransferases/metabolism , Apoptosis/drug effects , Epithelial Cells/drug effects , Fibroblasts/drug effects , Lung/drug effects , Microtubule Proteins/metabolism , Oligopeptides/pharmacology , Silicon Dioxide/toxicity , Silicosis/drug therapy , Transforming Growth Factor beta1/toxicity , A549 Cells , Animals , Collagen Type I/metabolism , DNA Breaks, Double-Stranded , Disease Models, Animal , Epithelial Cells/enzymology , Epithelial Cells/pathology , Fibroblasts/enzymology , Fibroblasts/pathology , Humans , Lung/enzymology , Lung/pathology , Male , Rats, Sprague-Dawley , Signal Transduction , Silicosis/enzymology , Silicosis/pathology , Up-RegulationABSTRACT
We performed liquid chromatography-tandem mass spectrometry (LC-MS/MS) on control and TGF-ß1-exposed rat lung fibroblasts to identify proteins differentially expressed between cell populations. A total of 196 proteins were found to be differentially expressed in response to TGF-ß1 treatment. Guided by these results, we next determined whether similar changes in protein expression were detectable in the rat lung after chronic exposure to silica dust. Of the five proteins selected for further analysis, we found that levels of all proteins were markedly increased in the silica-exposed rat lung, including the proteins for the very low density lipoprotein receptor (VLDLR) and the transmembrane (type I) heparin sulfate proteoglycan called syndecan 2 (SDC2). Because VLDLR and SDC2 have not, to our knowledge, been previously linked to the pathobiology of silicosis, we next examined whether knockdown of either gene altered responses to TGF-ß1 in MRC-5 lung fibroblasts. Interestingly, we found knockdown of either VLDLR or SDC2 dramatically reduced collagen production to TGF-ß1, suggesting that both proteins might play a novel role in myofibroblast biology and pathogenesis of silica-induced pulmonary fibrosis. In summary, our findings suggest that performing LC-MS/MS on TGF-ß1 stimulated lung fibroblasts can uncover novel molecular targets of activated myofibroblasts in silica-exposed lung.
Subject(s)
Fibroblasts/metabolism , Silicosis/genetics , Transcriptome , Transforming Growth Factor beta/pharmacology , Animals , Cells, Cultured , Collagen/genetics , Collagen/metabolism , Fibroblasts/drug effects , Male , Rats , Rats, Wistar , Receptors, LDL/genetics , Receptors, LDL/metabolism , Silicosis/metabolism , Syndecan-2/genetics , Syndecan-2/metabolismABSTRACT
Objective To reduce the perioperative complications of simultaneous pancreas and kidney transplantation (SPK ) and boost the clinical efficacy by exploring its perioperative management in diabetics with end-stage renal disease .Methods Retrospective analysis was performed for the clinical data of 5 diabetics with end-stage renal disease undergoing SPK since 2017 .Results The cold ischemic time of all allografts was under 8 hours .No surgical complications occurred in recipients post-transplantation .Two patients were complicated with hyperkalemia and their serum potassium levels normalized after treatment .One case of bleeding in abdominal cavity was cured conservatively . Renal functions of two patients with delayed renal function post-transplantation gradually recovered after hematodialysis . One case of peritransplant fluid collection recovered after debridement and drainage .Another case of acute left heart failure and cardiac arrest at 45 days post-transplantation resumed normal heart rhythm after rescue .However ,his consciousness could not be restored and his families gave up subsequent treatments . Transplanted kidneys and pancreases of these 4 patients normalized .Follow-up was conducted until March 20 , 2019 . They became insulin and dialysis independent and serum creatinine and blood glucose normalized .Diabetic complications were relieved and their quality-of-life also improved significantly .Conclusions SPK is an effective treatment for diabetics with end-stage renal disease .While maintaining normal serum creatinine and blood glucose , it may liberate patients from insulin dosing and dialytic maintenance ,lower diabetic complications and improve quality-of-life .
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Silicosis is the most common occupational lung disease in China, and is associated with a variety of complications, many of which are poorly understood. For example, recent data indicate that silicosis associates with the development of osteopenia, and in some cases this bone loss is severe, meeting criteria for osteoporosis. Although many factors are likely to contribute to this relationship, including a sedentary lifestyle in patients with advanced silicotic lung disease, we hypothesized that silica might directly reduce bone mineral density. In the present study, six Wistar rats were exposed to silica for 24â¯weeks in order to induce pulmonary silicosis and examine the relationship to bone mineral density. As expected, all rats exposed to silica developed severe pulmonary fibrosis, as manifested by the formation of innumerable silicotic nodules and the deposition of large amounts of interstitial collagen. Moreover, micro-CT results showed that bone mineral density (BMD) was also significantly reduced in rats exposed to silica when compared control animals and this associated with a modest reduction in serum calcium and 25-hydroxyvitamin D levels. In addition, we found that decreased BMD was also linked to increased osteoclast activity as well as fibrosis-like changes, and to the deposition of silica within bone marrow. In summary, our findings support the hypothesis that silicosis reduces bone mineral density and provide support for ongoing investigations into the mechanisms causing osteopenia in silicosis patients.
Subject(s)
Bone Density , Femur/pathology , Lung/pathology , Osteoporosis/chemically induced , Pulmonary Fibrosis/chemically induced , Silicon Dioxide , Silicosis/etiology , Tibia/pathology , Animals , Calcium/blood , Collagen/metabolism , Disease Models, Animal , Femur/diagnostic imaging , Lung/metabolism , Male , Osteoclasts/drug effects , Osteoclasts/metabolism , Osteoclasts/pathology , Osteoporosis/blood , Osteoporosis/diagnostic imaging , Osteoporosis/pathology , Pulmonary Fibrosis/metabolism , Pulmonary Fibrosis/pathology , Rats, Wistar , Risk Assessment , Severity of Illness Index , Silicosis/metabolism , Silicosis/pathology , Tibia/diagnostic imaging , Time Factors , Vitamin D/analogs & derivatives , Vitamin D/blood , X-Ray MicrotomographyABSTRACT
Objective To explore the regulation and mechanism of Cav1.2 current by KCNE1. Methods Transient transfection was used to transfer Cav1.2 channel plasmids separately or together with KCNE1 plasmids into HEK293 cells. The experiment was divided into 2 groups (15 cells in each group):Cav1.2 group, Cav1.2+KCNE1 group.The whole-cell patch clamp technique was used to record Cav1.2 current and gating dynamics. Results After co-transfection of KCNE1 with Cav1.2, Cav1.2 current decreased significantly. At 0 mV, peak current density of Cav1.2 was reduced from (-14.8±2.5) pA/pF to (-7.5±1.6) pA/pF (n=15, P<0.01). Based on the gate control mechanism, it is found that the regulation of Cav1.2 current by KCNE1 mainly makes the steady-state inactivation curve of the channel shifted to a more negative direction, thus accelerating the inactivation. Meanwhile, the recovery process of the channel after inactivation is slowed down and the recovery time constant was prolonged. Conclusions The KCNE1 subunit can reduce the current density of Cav1.2 by changing the channel inactivation and recovery process.
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Objective To observe the effect of allicin on the action potential duration (APD) and L-type calcium current (ICa,L) in the ventricular myocytes of rabbits with heart failure in order to explore the mechanisms of therapeutic effect of allicin on cardiac arrhythmias complicated with heart failure.Methods Forty-five New Zealand White male rabbits were randomly (random number) assigned to 3 groups (n=15 in each group):sham operated group (sham group),heart failure group (HF group),and heart failure treated with allicin group (HF+All group).The rabbit heart failure model was established by abdominal aortic constriction coupled with aortic regurgitation,the ventricular myocytes were obtained by enzyme double digestion,and the whole cell clamp was used to record action potential and calcium current.The action potential duration (APD),Ica,L and gating mechanism were observed during heart failure and allicin administered.Data were processed with pCLAMP version 10.2.Statistical analysis was performed using SPSS 17.0.Comparisons among groups were carried out using ANOVA,and SNK-q was used for multiple comparison as post-hoe.Results (1) Prolonged APD was found during heart failure,APD50 was prolonged from (93.4±4.7) ms in sham group to (115.5±6.2) ms in HF group(P<0.01).After administration of allicin 30 μmol/L,APD50 was shortened to (105.2±5.5) ms (P<0.05).(2) The density of ICa.L increased during heart failure,peak current density increased increased from (-8.4±0.6) pA/pF in sham group to (-15.1± 1.1) pA/pF while 0 mV attained at depolarizations (P<0.01).After administration of allicin 30 μmol/L,the current density reduced to (-10.1+0.8) pA/pF (P<0.01).The effect of allicin presented in both voltage dependent and consentration dependent manner.(3) According to the gating mechanism study,the main mechanism of lowering the density of ICa,L by allicin after heart failure was the acceleration of the steady inactivation of the channel,and the de-escalation of the recovery kinetic after the inactivation of the channel.Conclusions Allcin can be used to reduce the calcium current of ventricular myocytes in animal heart failure model,it has the potential of clinical use in treating cardiac arrhythmias during heart failure.
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Objective To observe the effect of KN93, a CaMK Ⅱ inhibitor, on delayed afterdepolarization (DAD) and calcium ion in ventricular myocytes of rabbits with heart failure, and to investigate the effect of CaMK Ⅱ signaling pathway on trigged arrhythmia after heart failure. Methods Thirty male New Zealand White rabbits were randomized(random number) into the sham operated group (sham group), heart failure group (HF group) and heart failure with KN93 group (HF+KN93 group) (n=10 each group). The rabbit heart failure model was established by abdominal aortic constriction combined with aortic valve regurgitation. The ventricular myocytes were isolated by double enzyme digestion. The action potential and the transient inward current (Iti) were recorded by the whole-cell patch-clamp. The intracellular calcium transient was measured by the ion concentration measurement system. The main calcium transporter protein was detected by Western blotting. Data were analyzed by pCLAMP10.2. Statistical analysis was performed using SPSS 17.0. Comparisons among groups were conducted using ANOVA, and SNK-q multiple comparison procedure was utilized for post-hoc analysis.Results (1) After induction of heart failure, DAD and increment of trigger activity (TA) were observedin rabbit ventricular myocytes. Treatment of KN93 with 1.0 μmol/L reduced the events of DAD and TA.(2) After induction of heart failure, Iti densities were increased from -0.12±0.02 pA/pF to -0.95±0.06pA/pF at the polarization potential of -50 mV (n=10, P<0.01). The current densities were reduced to -0.44±0.04 pA/pF after application of 1.0 μmol/L of KN93 (n=10, P<0.01). (3) KN93 led to decrementof intracellular calcium ion concentration and calcium transient amplitude, and acceleration of the decayprocess of calcium transient. (4) KN93 upregulated the expression of pPLN and SERCA2a, increased the uptake of intracellular calcium ion, downregulated the expression of NCX, decreased the Iti, and reduced the occurrence of DAD and TA. Conclusions KN93 can reduce the intracellular calcium ion concentration of the heart failure animal model, and the occurrence of the DAD and TA. CaMK Ⅱ may be a new therapeutic target for arrhythmias in the heart failure.
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OBJECTIVE: observe the changes of localization and expression of acetylated tubulin alpha( Ac-tubulin α) in fibroblasts of silicosis fibrosis. METHODS: i) A total of 22 autopsy case with coal workers' pneumoconiosis( CWP) was selected as CWP group( autopsy lesion lung tissue) and their adjacent normal lung tissues surrounding fibrotic regions were selected as the control group using typical sampling method. ii) Specific pathogens free healthy male rats were randomly divided into control group and 5 dust-exposed groups with 10 rats in each group. In dust-exposed group,the rats were exposed to 2 000 mg/m~3 of silica dust for 0,2,4,8,12 and 16 weeks respectively by dynamic inhalation. The rats in control group were given no treatment. iii) Primary cultured rat lung fibroblasts were induced by Ang Ⅱ at a concentration of 100 nmol/L at different time points( 0,5,15 and 30 min,1,3,6,12,24 and 48 hours). iv) The expression of Ac-tubulin α and α-smooth muscle actin( α-SMA) were detected by immunohistochemical staining. The expression of Ac-tubulinα/Vimentin and Ac-tubulin α/α-SMA were observed by immunofluorescence staining. The relative expression ofα-SMA and Ac-tubulin α protein was detected by Western blot. RESULTS: The positive expression of Ac-tubulin α in lung tissue of CWP group was down-regulated while that of α-SMA was up-regulated compared with the control group( P < 0. 01). Immunofluorescence results showed that Vimentin positive expression in the CWP group was significantly increased in the lesion fibrosis area,while Ac-tubulin α expression was absent. The co-expression of Ac-tubulin α and Vimentin was detected in normal lung tissue of control group. The animal experiment results showed that the expression of Ac-tubulin αin lung tissue of rats was down-regulated and that of α-SMA was up-regulated with the prolongation of dust exposure time( P < 0. 01). The Ang Ⅱ induced the differentiation of fibroblasts into myofibroblasts,and the expression of Ac-tubulin α in fibroblasts gradually decreased and the expression of α-SMA gradually increased with the prolongation of Ang Ⅱ induction time( P < 0. 05). CONCLUSION: The lack of expression of Ac-tubulin α may be involved in the occurrence and development of silicotic fibrosis.
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BACKGROUND:Heat Induction Typodont System (HITS) and Double-Slot Lingual Bracket System are patented products of the research group.It is very important to explore a good control of anterior tooth torque on the micro-implant-double-slot lingual bracket system.OBJECTIVE:To provide the experimental basis for force system applied for controlling the anterior tooth torque in lingual orthodontics through Typodont experiment based on the HITS.METHODS:Sixteen Class Ⅱ1 maxillary Typodont models without first premolar were made.Micro-implants were implanted in the lingual region of posterior teeth and labial region of anterior teeth.The direction of the retracting force was adjusted by changing the position of the lingual micro-implant (with a distance of 4,8 mm from the alveolar crest) and the length of the hook (4,8 mm).And lingual retracting force (150,300 g) and labial intruding force (50,100 g) were loaded.The tooth movement by HITS was simulated and the models were scanned before and after force loading.Then the three-dimensional images were reconstructed by Mimics 17.0.Factorial variance analysis was adopted to compare the anterior movement changes under different loading modes.RESULTS AND CONCLUSION:When the length of the hook was 4 mm/8 mm and the lingual micro-implant was 6 mm/10 mm from the alveolar crest,the displacement difference between the incisal edge and the root of the anterior teeth was smaller than other groups.The optimal mechanics was 150 g for the lingual retracting force and 100 g for the labial intruding force.It could provide a satisfactory control to the anterior teeth torque when retracting force and labial intruding force were loaded at the same time during the space closing phase of lingual treatment.This study based on HITS provided a scientific basis for the clinical application of micro-implant-double-slot lingual bracket system in space closing phase.
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Silicosis is the most serious occupational disease in China. The objective of this study was to screen various proteins related to mechanisms of the pathogenesis of silicosis underlying the anti-fibrotic effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) using proteomic profile analysis. We also aimed to explore a potential mechanism of acetylated α-tubulin (α-Ac-Tub) regulation by Ac-SDKP. Two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/TOF MS) were used to assess the different protein expression profiles between control and silicosis rats treated with or without Ac-SDKP. Twenty-nine proteins were identified to be potentially involved in the progression of silicosis and the anti-fibrotic effect of Ac-SDKP. Our current study finds that 1) the lost expression of Ac-Tub-α may be a new mechanism in rat silicosis; 2) treatment of silicotic rats with N-acetyl-Seryl-Aspartyl-Lysyl-Proline (Ac-SDKP) inhibits myofibroblast differentiation and collagen deposition accompanied by stabilizing the expression of α-Ac-Tub in vivo and in vitro, which is related with deacetylase family member 6 (HDAC6) and α-tubulin acetyl transferase (α-TAT1). Our data suggest that α-Ac-Tub regulation by Ac-SDKP may potentially be a new anti-fibrosis mechanism.
Subject(s)
Protein Processing, Post-Translational , Pulmonary Fibrosis/metabolism , Silicosis/metabolism , Tubulin/metabolism , Acetylation , Acetyltransferases/metabolism , Actins/metabolism , Amino Acid Sequence , Angiotensin II/physiology , Animals , Cells, Cultured , Fibroblasts/metabolism , Gene Ontology , Histone Deacetylase 6/metabolism , Lung/metabolism , Lung/pathology , Male , Protective Factors , Protein Stability , Pulmonary Fibrosis/chemically induced , Rats, Wistar , Silicon DioxideABSTRACT
<p><b>OBJECTIVE</b>To explore the inhibition effect and mechanism of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP)on myofibroblast differentiation via regulating acetylated tubulin α (Ac-Tub α)in vivo and in vitro.</p><p><b>METHODS</b>Silicotic model were made by SiO2 douched and divided into 6 groups as follows: control (4w, 8w)group, silicotic model (4w, 8w)group and post-or pre-treatment by Ac-SDKP group. Pulmonary fibroblasts were divided into 5 groups: (1) control; (2) Ang II; (3) Ang II+Ac-SDKP; (4) Ang II+Valsartan; (5) Ang II+TCS histone deacetylase (HDAC)6 20b. The localization of Ac-Tub α and α-smooth muscle actin (SMA) were observed by immunohistochemical (IHC) and immunofluorescence staining. The protein levels of Ac-Tub α, α-SMA, collagen type I (col I) and HDAC6 were measured by western blot.</p><p><b>RESULTS</b>In silicotic nodules and interstitial fibrosis area, positive expression of α-SMA, a classical marker of myofibroblast, was ob-served by IHC, accompanied with absence expression of Ac-Tub α. Furthermore, Ac-SDKP post-treatment could attenuate the levels of col I, α-SMA and HDAC6 to 48.39%, 52.63% and 70.18% compared with the silicotic 8w group respectively. And in Ac-SDKP pre-treatment group, compared with the silicotic 8w group, these protein levels were decreased to 32.26%, 64.91% and 54.39% respectively (P<0.05). The up-regulation of Ac-Tub α was found in Ac-SDKP post-and pre-treatment and increased to 3.00 and 2.90 folds compared with the silicotic 8w group. Compared with control group, the levels of α-SMA, HDAC6 and col I in Ang II group were up-regulated to 1.66, 3.56 and 4.00 folds accompanied with down-regulation of Ac-Tub by 44.44% (P<0.05). Pre-treatment with Valsartan, TCS HDAC6 20b or Ac-SDKP could inhibited all this changes induced by Ang II in vitro.</p><p><b>CONCLUSION</b>Ac-SDKP can inhibit the myofibroblast differentiation and collagen deposition via sup-press HDAC6 and up-regulate the expression of Ac-Tub α in vivo and in vitro.</p>
Subject(s)
Animals , Rats , Actins , Metabolism , Cell Differentiation , Collagen Type I , Metabolism , Disease Models, Animal , Fibroblasts , Cell Biology , Lung , Pathology , Myofibroblasts , Cell Biology , Oligopeptides , Pharmacology , Silicon Dioxide , Toxicity , Silicosis , Drug Therapy , Tubulin , MetabolismABSTRACT
[Abstract ] Objective Silicosis is one of the most serious occupational diseases in China .In this study,we explored the reg -ulatory effect of N-acetyl-seryl-aspartyl-lysyl-proline ( Ac-SDKP ) on angiotensin ( Ang ) Ⅱ-induced extracellular signal-regulated ki-nase ( ERK1/2) and Jun N-terminal kinase ( JNK) signals and its inhibitory effect on the differentiation of human embryonic lung MRC-5 fibroblasts to myofibroblasts via Ang Ⅱ-induced ERK1/2 and JNK signals . Methods Human embryonic lung MRC-5 fibro-blasts were induced by Ang Ⅱand pre-treated with the JNK signal inhibitor ( SP600125 ) , the ERK1/2 signal inhibitor ( PD98059 ) or Ac-SDKP.The proliferation of the cells was measured by MTT assay .The expressions of αS-MA, SRF, p-ERK1/2 and p-JNK were determined by immunocytochemical staining , and the expression levels of these proteins and collagen Ⅰwere detected by Western blot .Results The A value of Ang Ⅱ group (0.56 ±0.08) measured by MMT assay was 2.07 fold as control group ( 0.27 ±0.05 ). Pretreatment with SP600125 , PD98059 and Ac-SDKP, the A value were (0.39 ±0.02), (0.40 ±0.03) and (0.36 ±0 0.5) that had a statistical significance with Ang Ⅱgroup.The up-regulation of colla-gen type Ⅰ,α-SMA, SRF were induced by Ang Ⅱ by 4.50, 3.50 and 3.00 fold compared with control group.Moreover, the expression of p-ERK1/2 and p-JNK were increased as 6.71 and 7.90 fold as control. Pre-treatment with Ac-SDKP could inhibit p-JNK and p-ERK1/2 to 29.79% and 46.84% compared with AngⅡ group. Conclusion Ac -SDKP can inhibit the differentiation of human embryonic lung MRC-5 fibroblasts to myofibroblasts by regulating AngⅡ-induced JNK and ERK1/2 signals.
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AIM: To detect whether N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) inhibits epithelial-mes-enchymal transition in A549 cells induced by TGF-β1 through suppressing the expression of heat shock protein 27 (HSP27) and zinc finger proteins Snail (including SNAI1and SNAI2) which ultimately inhibited the deposition of type I and type III collagens.METHODS:The colocalizations of HSP27 and SNAI1/SNAI2 respectively on A549 alveolar epi-thelial cells induced by TGF-β1 were measured by confocal microscopy .The expression of HSP27, SNAI1 and SNAI2 at mRNA level was detected by real-time PCR.Western blotting analysis was used to detect the expression of HSP 27, SNAI1 and SNAI2 on epithelial-mesenchymal transition in A549 cells induced by TGF-β1 and also the deposition of type I and type III collagens in A549 cells transfected with HSP27shRNA prior to TGF-β1 stimulation.RESULTS: Compared with control group, TGF-β1 increased the expression of HSP27, SNAI1, SNAI2, type I and type III collagen, which decreased significantly followed by Ac-SDKP intervention.The expression of SNAI1, type I and type III collagen decreased signifi-cantly after transfected with HSP27shRNA in A549 cells, which had the similar effect on Ac-SDKP intervention.CON-CLUSION:Ac-SDKP inhibits the transition of cultured A 549 cells to myofibroblasts and attenuates collagen synthesis by suppressing the expression of HSP 27 and zinc finger proteins SNAI 1 and SNAI2.
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<p><b>OBJECTIVE</b>To perform a comparative proteomic analysis for identification of pulmonary proteins related to the progression of silicosis and anti-fibrotic effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP).</p><p><b>METHODS</b>Bronchial instillation of SiO₂powder (for 4 or 8 weeks) was applied in rats to establish a silicosis model. Ac-SDKP treatment was performed before (prevention group) or after (treatment group) SiO₂instillation. The control group was treated by bronchial instillation of sodium chloride solution of the same volume as SiO₂powder for 4 or 8 weeks. Proteins in lung tissue were separated by two-dimensional gel electrophoresis and stained with colloidal Coomassie brilliant blue. The gel images were scanned with the Lab Scan III system and analyzed with Imagemaster 6.0. The protein spots with significant differences between two groups (i.e., P value was less than 0.05 in One-way ANOVA) and with a change in volume over 30% were defined as differential proteins. Comparison was performed between the silicosis group and control group after 4 or 8 weeks, between the Ac-SDKP treatment group and silicosis group after 8 weeks, and between the Ac-SDKP prevention group and silicosis group after 8 weeks. The differentially expressed proteins were subjected to in-gel digestion with trypsin and MALDI-TOF-MS and Mascot search engine analysis to identify these proteins.</p><p><b>RESULTS</b>Thirty-three differential proteins were identified. In comparison with the control group (4 weeks), the silicosis group (4 weeks) had 17 up-regulated proteins and 11 down-regulated proteins. In comparison with the control group (8 weeks), the silicosis group (8 weeks) had 16 up-regulated proteins and 12 down-regulated proteins. In comparison with the silicosis group (8 weeks), the Ac-SDKP treatment group had 5 up-regulated proteins and 6 down-regulated proteins, and the Ac-SDKP prevention group had 8 up-regulated proteins and 10 down-regulated proteins.</p><p><b>CONCLUSION</b>Critical regulatory proteins related to silicotic fibrosis and anti-silicotic effect of Ac-SDKP have been identified. These proteins may play an important role in proliferation, apoptosis, inflammation, epithelial-mesenchymal transition, and signal transduction in silicosis.</p>
Subject(s)
Animals , Male , Rats , Disease Models, Animal , Lung , Metabolism , Oligopeptides , Therapeutic Uses , Proteome , Metabolism , Rats, Wistar , Silicosis , Drug Therapy , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the inhibition effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) on myofibroblast differentiation of MRC-5 human fetal lung fibroblasts induced by angiotensin (Ang) II.</p><p><b>METHODS</b>The study was divided into 2 step: (1) MRC-5 human fetal lung fibroblasts was induced for 48 h at different dose of Ang II and at different time point by 100 nmol/L Ang II. Then the expression of collagen type I and α-smooth muscle actin (α-SMA) were mesaured by western blot. (2) MRC-5 human fetal lung fibroblasts were divided into 4 group: (1) control, (2) Ang II, (3) Ang II+Ac-SDKP, (4) Ang II+8-Me-cAMP (a specific activator of Epac). The α-SMA expression was observed by immnocytochemical stain. The protein expression of collagen type I, α-SMA, serum response factor (SRF), myocardin-related transcription factor (MRTF)-A, exchange protein directly activated by cAMP (Epac) 1, 2 were measured by Westen blot.</p><p><b>RESULTS</b>Myofibroblast differentiation could be induced by Ang II from MRC-5 cells with a dose- and time-dependent manner. The up-regulation of SRF and MRTF-A were observed in MRC-5 cells induced by Ang II and accompanied with collagen I and α-SMA increased. Pre-treatment with 8-Me-cAMP or Ac-SDKP could attenuated all this changes induced by Ang II, and promoted the expression of Epac1.</p><p><b>CONCLUSION</b>Ac-SDKP can inhibit the myofibroblast differentiation of MRC-5 cells induced by Ang II via Epac1 activating.</p>
Subject(s)
Humans , Actins , Angiotensin II , Cell Differentiation , Collagen , Collagen Type I , Cyclic AMP , Fetus , Cell Biology , Fibroblasts , Cell Biology , Guanine Nucleotide Exchange Factors , Lung , Cell Biology , Myofibroblasts , Oligopeptides , Pharmacology , Serum Response Factor , Trans-ActivatorsABSTRACT
Objective To explore an operative method which can adequately enlarge the inner canthus with inconspicuous scar and perfect shape based on the transverse incision for epicanthus correction,and to evaluate the short-term results and long-term effects comprehensively.Methods The surface projection of tears port medial point was designed as a new inner canthal point,the full thickness skin from new inner canthal point was cut to the free edge of epicanthus,and an outward prolonged incision was made along the lower eyelid edge,separating the subcutanous dissociation or cutting off part of displaced orbicularis,so to relieve the skin tension of inner canthal and reset skin,suturing the new inner canthal point and lower eyelid edge.Results 120 cases were followed up for more than one year,the complication rate at different postoperative time points and patient satisfactory rate one year after operation were compared.12 cases had mild scar formation,from six months to one year after operation,the scar gradually became softened and then inconspicuous.22 cases had a little skin fold at the inner canthal in postoperational one month.2 cases were unsatisfied with the shape of inner canthal,Others were satisfied with therapeutic efficacy,the correction was sufficient with inconspicuous scar and 1ess recrudescence.Conclusions Epicanthus correction by subciliary incision epicanthoplasty is a simple and effective method and suitable for all types of epicanthic fold,and it has the advantages of sufficient enlarged inner canthus,inconspicuous scar and 1ess recrudescence.
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Objective To discuss the efficacy and advantage of dolabriform flap in repairing periorbital skin and soft tissue defects.Methods 21 patients with periorbital skin disease were treated by radical resection,and then the adjacent or distant dolabriform flaps were designed for the one stage repair of skin and soft tissue defects in these cases according to the defect location,size,shape and neighborous skin conditions.The largest defect size was 3.5 cm × 4.5 cm,The smallest defect size was 1.2 cm × 1.5 cm.The largest ratio of hatchet pedicle width to axe handle length reached 1 ∶ 5,and the largest ratio of hatchet pedicle width to hatchet edge length was 1 ∶ 4.Results All of these dolabriform flaps in 21 patients survived well with primary incision healing.18 cases had been followed up for 3 to 12 months,showing that there was no deformation found on eyelids,eyebrow and nose,the colour of these flaps were similiar to that of the normal skin,without being fat and clumsy,all scars were almost invisible,and the cosmetic results were satisfactory.Conclusions Larger diameter of skin and soft tissue defect in different periorbital parts could be repaired individually with dolabriform flap according to the characters of natural lines,and it is a simple and flexible method for repairing periorbital skin defects,and can achieve satisfactory functional and cosmetic results.
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Human hepatocyte transplantation to treat liver-based metabolic deficiencies and acute liver failure has shown promising early improvement in liver function; however, long-term success has not been achieved. Stem cell transplantation to restore liver function as an alternative to whole liver transplantation has not been successful in humans. As alternative sources of cells for human hepatocyte transplantation, stem cells are under investigation. The liver extracellular matrix presents an ideal scaffold for stem cell differentiation into hepatocytes, as well as cell transplantation. The innovative technique of the decellularized liver matrix presents great potential as the scaffold for hepatocyte maturation and transplantation, and allows the development of engineered recellularized liver graft for transplantation.