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Objective:To observe the lipid-lowering effect of atorvastatin on patients with acute cerebral infarction with different ATP-binding cassette subfamily B member 1(ABCB1) genotypes, and thus to provide clinical research evidence for individual application of atorvastatin in patients with acute cerebral infarction.Methods:From March 2021 to December 2021, 131 patients with acute cerebral infarction admitted to the Department of Neurology of Xuchang Central Hospital were included. The ABCB1 G2677T gene polymorphism rs2032582 of patients was detected by fluorescence staining in situ hybridization.Based on the detection results, patients were divided into GG group, GT group and TT group.All patients were given atorvastatin (20 mg/d) for lipid-lowering treatment.The levels of low density lipoprotein cholesterol(HDL-C), high density lipoprotein cholesterol(HDL-C), total cholesterol(TC)and triglyceride(TG) in serum of patients in the three groups before and 2 months after treatment were recorded and analyzed.The adverse drug reactions in the three groups were recorded. When the serum LDL-C level was less than 1.8 mmol/L, it was considered that the lipid-lowering treatment was effective.The binary Logistic regression analysis was used to explore the influencing factors of atorvastatin lipid lowering therapy.The software of SPSS 25.0 was used for statistical analysis.Results:There were 50 (38.17%), 49 (37.40%) and 32 (24.43%) patients in GG group, GT group and TT group, respectively. The serum TC levels of patients in GG group, GT group and TT group after treatment were (3.47±0.70) mmol/L, (3.59±1.09) mmol/L and (3.48±1.02) mmol/L, respectively, which were lower than those before treatment ((4.27± 0.99) mmol/L, (4.02±0.98) mmol/L and (4.03±1.31) mmol/L), all of which were statistically significant ( t=7.652, 3.092, 5.593, all P<0.01). The serum LDL-C levels in GG group, GT group and TT group after treatment were (1.89±0.53) mmol/L, (2.07±0.92) mmol/L and (1.96±0.79) mmol/L, respectively, which were lower than those before treatment ((2.87±0.92) mmol/L, (2.56±0.89) mmol/L and (2.55±1.11) mmol/L) ( t=9.896, 4.055, 5.980, all P<0.001). The differences of serum LDL-C level before and after treatment in GG group, GT group and TT group were (-0.97±0.69) mmol/L, (-0.50±0.86) mmol/L and (-0.59±0.56) mmol/L, respectively. The difference of serum LDL-C level before and after treatment in the three groups was statistically significant ( F=5.614, P=0.005). The difference of TC, TG and HDL-C before and after treatment was not statistically significant( F=2.783, 0.490, 1.677, all P>0.05). The binary Logistic regression analysis showed that ABCB1 G2677T gene type and staying up late were independent influencing factors for atorvastatin lipid-lowering therapy. The probability of effective lipid-lowering in GT patients with ABCB1 G2677T gene was 27.9% of that in GG patients ( OR=0.279, 95% CI: 0.110-0.709, P=0.007), and the probability of TT type patients was 33.8% of GG type patients ( OR=0.338, 95% CI: 0.121-0.943, P=0.038). The probability of effective lipid-lowering in patients who had the habit of staying up late was 26.4% of the patients who did not stay up late ( OR=0.264, 95% CI: 0.118-0.591, P=0.001). There was no significant difference in the total incidence of adverse drug reactions among the three groups( χ2=0.868, P=0.648). Conclusion:The lipid-lowering effect in patients with GG type of ABCB1 G2677T is better than that of GT type and TT type when atorvastatin is used to treat patients with acute cerebral infarction.
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OBJECTIVE： To investigate the effects of rosuvastatin on in-stent restenosis in middle-aged patients with acute coronary syndrome（ACS）after percutaneous coronary intervention （PCI）. METHODS： Totally 400 middle-aged ACS patients underwent PCI were selected from Xuchang Central Hospital during Mar. 2016 to Apr. 2017， and then divided into control group and observation group according to random number table， with 200 patients in each group. Both groups were given conventional drugs for secondary prevention of coronary heart disease. Control group were given Clopidogrel hydrogen sulfate tablets 75 mg， once a day+Aspirin enteric-coated tablets 100 mg， once a day+Atorvastatin calcium tablets 20 mg， once at bed time every day orally after PCI. Observation group was given Clopidogrel hydrogen sulfate tablets 75 mg， once a day+Aspirin enteric-coated tablets 100 mg， once a day+Rosuvastatin calcium tablets 10 mg， once at bed time every day orally after PCI. Both groups were treated for consecutive 12 months. The serum levels of TG， TC， LDL-C， hs-CRP and IL-35 were recorded in 2 groups before surgery， 1， 3， 6 and 12 months after surgery； in-stent minimum lumen diameter （MLD） was observed immediately after surgery and 12 months after surgery. The occurrence of in-stent restenosis， major adverse cardiovascular events （MACE） and adverse drug reaction （ADR） were recorded. RESULTS： Totally 14 patients dropped out from control group and 18 from observation group， and 368 patients completed the study. Before surgery and immediately after surgery， there was no statistical significance in the serum levels of TG， TC， LDL-C， hs-CRP IL-35 or MLD （P＞0.05）. One day after surgery， the levels of hs-CRP were increased significantly in 2 groups， compared with before surgery （P＜0.05）. One， three， six and twelve months after surgery， the serum levels of TG， TC， LDL-C and hs-CRP were decreased significantly in 2 groups， while the levels of IL-35 were increased significantly， compared with before surgery； and the serum levels of TG and TC （1 and 3 months after surgery）， LDL-C （3 and 6 months after surgery） and hs-CRP （1 month after surgery） in observation group were significantly lower than control group； the level of IL-35 in observation group （1 month after surgery） was significantly higher than control group （P＜0.05）. Twelve months after surgery， MLDs of 2 groups were decreased significantly， and observation group was significantly higher than control group （P＜0.05）. There was no statistical significance in the incidence of in-stent restenosis or the total incidence of MACE and ADR between 2 groups after surgery （P＞0.05）. CONCLUSIONS： Rosuvastatin can effectively improve the levels of blood lipid and inflammatory factor in meddle-aged patients with ACS after PCI， and its effect is better than that of atorvastatin. The drug can delay in-stent restenosis after PCI in these patients， which is better than the effect of atorvastatin. At the same time， rosuvastatin can not increase the risk of MACE and ADR with good safety.
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Objective To investigate the effects of simulated microgravity by RCCS on proliferation and cell cytoskeleton of human HaCaT keratinocyte. Methods The rotary cell culture system (RCCS) was used to simulate the microgravity environment, and human HaCaT keratinocytes were divided randomly(random number) into the simulated microgravity group (SMG) and normal gravity group (NG). HaCaT cells in the two groups were harvested respectively after 32, 36 and 42 h culture. The HaCaT cells proliferation and cycles were detected by flow cytometry, the concentration of hb-EGF in supernatant was detected by ELISA, and the cell cytoskeleton was observed after 42 hours' culture under laser confocal microscope with FITC-labeled technique. SPSS 23.0 statistical software was used for statistical analysis, and P <0.05 was considered statistically significant. Results The flow cytometry showed that the proportions of human HaCaT keratinocytes in G1 and G2/M phases were increased while the proportion of HaCaT cells in S stage was decreased significantly after 32, 36 and 42 h RCCSculture compared with those in the normal gravity group. The HaCaT cells in G1 stage were declined along with incubation time. ELISA results showed that the hb-EGF concentration in HaCaT supernatant under simulated microgravity culture for 24 and 36 h was lower than that in the normal control group (P<0.01). The laser confocal microscope revealed that the HaCaT fluorescence intensity was decreased,and there were disordered microfilaments, structural ambiguity, pseudopodia reduction and irregularshape among FITC-labeled HaCaT cells cultured 42 h in RSSC compared with the normal gravity group.Conclusions RCCS simulated microgravity environment could inhibit the cell cycle transformation and proliferation of human HaCaT keratinocyte, affect the keratinocyte-secreting function, and induce alterations of the cell cytoskeleton.
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The rotary cell culture system(RCCS)was used to simulate the microgravity environment, and FRTL-5 cells were divided into simulated microgravity group(SMG)and normal gravity group(NG). FRTL-5 cells were harvested after treatment for 6, 12, 24, and 36 h, the cell viability was measured by MTT assay, and the cells cycles were detected by flow cytometry. The ultrastructure of FRTL-5 cells was observed under laser confocal microscope with FITC-labeled technique. The MTT assay showed that the proliferation of FRTL-5 cells was significantly inhibited after RCCS treatment for 6, 12, 24, and 36h compared with NG(P<0.05), in which the most obvious effect was observed at 24h. The flow cytometry showed that the proportion of FRTL-5 cells at G1 stage in RCCS group was increased significantly after 6, 12, 24, and 36h compared with NG(P<0.05), while the proportion of FRTL-5 cells at S stage was decreased significantly(P<0.05)except that cultured with RCCS for 6 h. The proportion of FRTL-5 cells at G2/M stage was decreased in early phase(6-12 hours)of RCCS culture, with the lowest at 12h and transient increase at 24h of RCCS culture. The laser confocal microscope revealed that there were local microfilament depolymerization, tension fibers decrease, structure disorder, cellular pseudopodia reduction, and irregular shape among FITC-labeled FRTL-5 cells cultured with RCCS for 36h. (Chin J Endocrinol Metab, 2018, 34: 598-601)
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BACKGROUND:The cell-sheet technology, based on a temperature-responsive culture, has been drawing more and more attention;however, the temperature-responsive culture dish is quite expensive. Therefore, it is imperative to develop a substitutive technique.OBJECTIVE: To study the feasibility of cell-sheet ulturing using common culture dish, and investigate the chondrogenesis of the cell sheet. METHODS: A piece of nasal septal cartilage was adopted from a patient with deviation of nasal septum to extract primary chondrocytes that were then cultured and amplified. The passage 3 chondrocytes were used to construct ell sheets. Monolayer cell sheet was formed by intensive culturing and allowing the extracellular matrix secretion. Bilayer cell sheet was constructed by seeding passage 2 chondrocytes on the monolayer cell sheet. The cell sheets were harvested using cell scraper, their properties were investigated prior to plantation into nude mice to construct the tissue-engineered cartilage. RESULTS AND CONCLUSION: Both bilayer and monolayer cell sheets with soft tremellose structures showed no significant difference through naked eyes. The newly harvested cell sheets appeared to have good fluidity and gelation. Eight weeks after mplantation into the nude mice, mature cartilage blocks were obtained. Histologically, the cell sheets were thin films composed by layered chondrocytes and extracellular matrix. Glycosaminoglycan formation and type Ⅱ collagen expressions were observed in the cell sheets cultured in vitro. The explanted samples exhibited ature cartilaginous tissue at 8 weeks after implantation. Biochemical analysis showed that the DNA contents of the neocartilages were higher than those of native human costal cartilage, while the contents of glycosaminoglycan and hydroxyproline were similar to native human nasal septal cartilage. To conclude, the hondrocyte cell sheets are likely to be constructed and harvested successfully using common culture dish, and the cell sheets exhibit favourable chondrogenesis.
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Objective]To study the feasibility of Cartilage engineering using fibrin gel and chondrocyte cell sheets.[Methods]rabbit auricular chondrocytes were isolated and cultured to form cell sheets in flasks. The cell sheets were harvested using cell scrapers,and cut into fragments. The two precursor solutions of Fibrin gel were used to suspend the cell sheet fragments and isolated chondrocytes,and then added into the wells of a 48-well plate to form Gelatinous chondroid disc constructs. After in vitro culture, the constructs were implanted into nude mice. After 8 weeks,the constructs were harvested,and the specimens were evaluated using grossly observing, histological and immunohistochemical observation. [Results]Mature cartilage discs were obtained. The histomorphology of the explanted discs appeared non-uniform cartilaginous tissue comprise of regenerated cartilage islands with different size and irregular shape. Immunohistochemistry staining demonstrated that type II collagen highly expressed in the ECM of the cartilage islands. In 1 of the 8 discs,partial ossification was observed.[Conclusion]Fibrin gel is a favourable carrier. Artificial cartilage with stereochemical structure was constructed via combining the fibrin gel and chondrocyte cell sheets.
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Objective To investigate the effects of simulated microgravity by rotary cell culture system (RCCS) on expression profiles of miRNA in mouse fibroblasts L929 cell line.Methods L929 cells were cultured in vitro and divided into simulated microgravity (SMG) group and normal gravity (NG) group according to the simple random method.Samples of two groups were collected on 7th day of culture and the total RNAs were extracted,labeled,and hybridized in sequence.Feature Extraction Software was used to collect the array images and get raw data,which were analyzed by Genespring Software.Differentially expressed miRNAs were identified and then validated by qRT-PCR.Target genes of differentially expressed miRNAs were predicted by the databases of Targetscan and microRNAorg,and the intersections of databases were identified as potential regulatory target genes.Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis were applied to determine the roles of these target genes.Relevant biological functions and/or signaling pathways of the regulated genes especially related with wound healing process were categorized based on their enrichments.Then integration predictions of the miRNA and mRNA expression profiles had been proposed to refine the functional miRNA-mRNA relationships.The miRNA-mRNA functional network and miRNA-mRNA control network were constructed.Results Four miRNA genes were up-regulated significantly including mmumiR-669j,-122-5p,-30a-3p,-6516-3p,among which mmu-miR-669j was up-regulated at 52.84 folds with the greatest significance (P ＜ 0.05).Seventeen miRNA genes were down-regulated significantly including mmu-miR-21a-3p,-miR-28a-5p,-218-5p,-210-3p,-miR-19a-3p,-miR-31-3p,and-miR-19b-3p,among which mmu-miR-28a-5p was down-regulated at 15.47 folds with the greatest significance (P ＜ 0.05).The qRT-PCR showed a high concordance with the microarray results (P ＜ 0.05).Target gene prediction and functional enrichment analysis suggested that a variety of biological processes and signaling pathways involved in wound repair were significantly enriched (P ＜ 0.05).Function network and regulation network of miRNA-mRNA covered all the differentially expressed miRNAs,which suggested that miR-21 a-3p and predicted target gene Smad3 might play an important role in wound healing under microgravity.Conclusions Simulated microgravity by RCCS can significantly affect the expression of stress-related miRNAs in mouse fibroblasts L929.The miRNA target gene prediction and functional enrichment analysis based on gene chip technology may provide theoretical basis for illustrating the mechanism and management of weightlessness stress injury.
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BACKGROUND:To seek for ideal scaffold materials is still an important task for cartilage tissue engineering.OBJECTIVE:To investigate the application of the AviteneTM microfibrillar collagen hemostat sponge in cartilage tissue engineering.METHODS:Rabbit auricular cartilage was harvested via surgical operation,and primary chondrocytes were isolated and amplified.Microfibrillar collagen hemostat sponge was cut into small bricks.The passage 2 chondrocytes were suspended and seeded onto the spongy bricks.After 1 week of in vitro culture,the constructs were then implanted into nude mice.After 8 weeks,the specimens were collected and evaluated using gross,histological and immunohistochamical observation.RESULTS AND CONCLUSION:During the cell seeding,the scaffold maintained its dimensions.No shrinkage was observed when the cell suspension was added.There was no considerable change in dimensions during the 1-week in vitro culture and at 8 weeks after implantation in nude mice.At 8 weeks post-implantation,mature cartilage blocks were harvested,which were white,translucent,and flexible.Histologically,the constructs appeared to have typical mature cartilaginous tissues,with robust extracellular matrix secretion,in which the microfibrillar collagen was incompletely degraded.We conclude that the microfibrillar collagen is a favorable scaffold material for cartilage tissue engineering.
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Objective@#To explore the utility of pharyngeal pH monitoring which positive standard is Ryan index in diagnosis of laryngopharyngeal reflux disease.@*Methods@#In a retrospective study, clinical data of 590 patients who had symptoms laryngopharyngeal reflux disease from February 2016 to March 2017 were analyzed. All patients were received electronic laryngoscopy, assessment of reflux symptom index(RSI) and reflux finding score(RFS), and pharyngeal pH monitoring. SPSS 19.0 software was used to analyze the date.@*Results@#There were 94 patients whose Ryan index were positive(15.93%). Among the 94 patients, 70 were positive during upright, 12 during supine and 12 during both upright and supine. There were 40 patients(6.78%)with pH decline events related to symptoms, while those Ryan index were normal. There were 536(90.85%), 417(70.68%), 233(39.49%) and 117(19.83%) patients with pH<6.5, pH<6.0, pH<5.5 and pH<5.0 events respectively. The positive rate of RSI, RFS, RSI and RFS, RSI or RFS were 44.24%, 16.78%, 7.12%, 53.90% respectively. The RFS score in Ryan index positive group was higher than that in Ryan index negative group[(8.2±2.4) vs (4.0±2.9), u=5.424, P<0.05], while the RSI score in Ryan index positive group was not statistically different from that in Ryan index negative group[(11.3±6.2) vs (12.7±5.8), t=1.247, P=0.167].@*Conclusions@#Pharyngeal pH monitoring is an objective and non-invasive method which can reflect laryngopharyngeal reflux directly. However, with the Ryan index as a criterion for the diagnosis of laryngopharyngeal reflux disease, partial patients may be missed. Further studies are needed to obtain more accurate and objective laryngopharyngeal pH statistical index for diagnosis of laryngopharyngeal reflux disease.
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Objective To investigate the animal model of acute lung injury (ALI) induced by caudal vein injection of trypsin in rats and to evaluate the model.Methods The model of lung injury was established by caudal vein injection of trypsin in rats.The rats were killed at the time point of 3 h,6 h,12 h,24 h,and 24 h and then the pathological changes of structure of lungs,peripheral blood neutrophil count,arterial blood gas analysis and lung wet/dry (W/D) weight ratio in rats were measured and observed.Results The results of hematoxylin eosin (HE) staining showed that there was no obvious pathological changes in lung tissues of the control group,while alveolar and pulmonary septal edema,thickening,and a large number of inflammatory cells infiltration in the lung tissues of the model group.Compared to the control group,the peripheral blood neutrophil counts,W/D and PaCO2 were significantly increased,PaO2 was significantly decreased (P ＜0.01).There was significant differences in the number of peripheral blood neutrophils,PaCO2,W/D and PaO2 between the model groups (P ＜ 0.01).Conclusions The rat model of ALI induced by trypsin can successfully simulate the lung damage caused by the release of a large number of trypsin when severe acute pancreatitis occurred.
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Objective To investigate the effect of simulated microgravity on growth , morphology, protein expression and virulence gene expression of Klebsiella pneumoniae (KPN).Methods KPN was divided into simulated microgravity group and control group in the experiment .The former group was in the ambient of simulated microgravity in a clinostat .The bacterial growth curves , morphologyical changes in electron microscopy , and protein expression were detected by SELDI-TOF-MS, and the expression of 4 virulence genes(ureA,wabG,uge and fimH) by real-time fluorescence quantitative PCR (RT-PCR) in both groups.Results Compared with the control group , the growth of KPN under simulated microgravity was accelerated , and the total bacterial count increased in microgravity group .The bacterial morphology in microgravity group was changed under scanning electron microscopy (SEM), and thinner and longer bacteria were increased .The transmission electron microscopy ( TEM) analysis revealed increase in cytoplasmic granular substance in microgravity group .Proteome analysis showed that the expression of 18 proteins was changed , half of which up-regulated and the rest were down-regula-ted.Those 18 proteins were searched in the protein library .And 21 proteins of a similar molecular mass were retrieved ,13 of which,proteins with known functions ,were closely related to bacterial life activities .RT-PCR results showed that four virulence genes of KPN were down-regulated.Conclusion Upon exposure to simulated microgravity , the growth and repro-duction of KPN are accelerated and enhanced .The bacterial morphology is changed .The strain′s protein expression and four virulence genes expressionare also changed .Therefore,microgravity can change the characteristics of KPN .
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<p><b>OBJECTIVE</b>To detect the number of cells and the level of IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-γ and IL-17 cytokines in the peripheral blood of mice exposed to rocket kerosene by skin.</p><p><b>METHOD</b>ICR mice were randomly divided into the normal control group and RK experimental group (400 µl×1 group). RK undiluted fuel were applied directly to the dorsal skin of the mice. In control groups were treated with sesame oil (SO). the number of blood cells were detected by automatic blood cell counter and the level of IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-γ and IL-17 cytokines in serum were detected by using flow cytometry and BD CBA Flex set kit.</p><p><b>RESULT</b>Compared with the normal group, WBC and LYM had a decreasing tendency 2 h and decreased significantly 6 h, 12 h and 1 d after RK exposure (P<0.05). They increased significantly 7 d after RK exposure (P<0.05). Compared with the normal group, the level of IL-6 increased significantly 2 h, 6 h, 12 h,1 d and 3 d (P<0.05). The level of TNF-α increased significantly 2h, 3d, 5d and 7d (P<0.05). The level of IL-10 increased significantly 2 h, 6 h, 3 d, 5 d and 7 d (P<0.05). The level of IFN-γ increased significantly 6 h and 3 d (P< 0.05). The level of IL-17 significantly increased 3 d, 5 d and 7d (P<0.05).</p><p><b>CONCLUSION</b>RK can change the number of immune cells, causing the immune cytokine changes in mice after RK cutaneous exposure.</p>
Asunto(s)
Animales , Ratones , Administración Cutánea , Recuento de Células Sanguíneas , Citocinas , Sangre , Citometría de Flujo , Queroseno , Toxicidad , Ratones Endogámicos ICRRESUMEN
Objective To study the effect of osgentide (OST) on proliferation of mouse preosteoblast MC3T3-E1 under simulated microgravity ( SMG ) .Methods Under normal conditions , cell proliferation was evaluated by MTT assay to screen an OST compound of an effective concentration after MC 3T3-E1 cells were treated with series OSTs .Furthermore, cell proliferation and cell cycle distribution of MC 3T3-E1 cells were analyzed after treatment with 1 nmol/L OST5 by MTT assay and by flow cytometry ( FCM) scanning under SMG .Results Under normal conditions , 1 nmol/L OST5 was able to significantly promote the proliferation of MC3T3-E1 cells (P<0.01).Under SMG, proliferation of MC3T3-E1 cells was significantly inhibited and more cells entered G 1 than under normal conditions (CN).The proportion of S phase of MC3T3-E1 cells after treatment with 1 nmol/L OST5 ( OST-SMG) for 3 d was higher than that of untreated MC 3T3-E1 cells under SMG,suggesting that OST5 could promote DNA synthesis ( P<0.05 ) .Conclusion OST5 facilitates the proliferation of MC3T3-E1 cells under SMG, which provides a basis for the use of OST5 in the prevention and treatment of bone loss relat-ed to microgravity .