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ObjectiveTo observe the modulatory effect of modified Zhenwutang on the interleukin-6 （IL-6）， matrix metallopeptidase-9（MMP-9）， type Ⅳ collagen（COL-Ⅳ） in rats with chronic renal failure （CRF） and to investigate the potential mechanism of its treatment of CRF. MethodFifty male SD rats were randomly divided into a modeling group of 40 rats and a normal group of 10 rats， and the modeling group was prepared by continuous adenine gavage for 12 weeks. After successful modelling， the modelling group was divided into the model group， the low dose （7.2 g·kg-1·d-1） group， the medium dose （14.4 g·kg-1·d-1） group， the high dose （28.8 g·kg-1·d-1） group and the Benadryl hydrochloride （10 mg·kg-1·d-1） group for gavage according to the random number table method， In the normal group and the model group， equal volume of distilled water was administered by gavage for 4 weeks. After the administration， the levels of blood creatinine （SCr）， blood urea nitrogen （BUN） and 24 h urine protein （24 h-UTP） were measured， the levels of serum IL-6 were measured by enzyme linked immunosorbent assay（ELISA）. Immunohistochemistry （IHC） was used to detect intercellular cell adhesion molecule-1 （ICAM-1）， IL-6， MMP-9， and other molecules in the rat kidney. The expression of ICAM-1 mRNA， IL-6 mRNA， MMP-9 mRNA and COL-Ⅳ mRNA in rat kidney tissues was measured by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression levels of ICAM-1， IL-6， MMP-9 and COL-Ⅳ in rat kidney tissues were measured by Western blot. ResultCompared with the normal group， the levels of SCr， BUN and 24 h-UTP were significantly increased in the model group （P<0.01）； the serum IL-6 level was significantly increased （P<0.01）， the tubular lumen was dilated with atrophy， the tubular epithelial cells were necrotic， swollen and vacuolated， the interstitium was infiltrated by a large number of inflammatory cells and collagen fibers were deposited， the levels of IL-6， ICAM-1 and COL-Ⅳ were strongly positive in the tubular interstitium of the model group （P<0.01）， The levels of ICAM-1 mRNA， IL-6 mRNA and COL-Ⅳ mRNA were significantly increased （P<0.01） and MMP-9 mRNA was significantly decreased （P<0.05） in the model rats. ICAM-1， IL-6 and COL-Ⅳ protein expression was significantly increased （P<0.01） and MMP-9 protein expression was significantly increased （P<0.01） in the renal tissue， and MMP-9 protein expression was significantly decreased （P<0.01）. Compared with the model group， the 24 h-UTP， SCr and BUN levels of rats were significantly reduced after treatment with modified Zhenwutang （P<0.01）， the serum IL-6 level was significantly decreased （P<0.01）， the renal lesions of rats were significantly improved and collagen fiber deposition was reduced； the expression of IL-6， ICAM-1 and COL-Ⅳ in renal tubules and interstitium was weakened， and MMP-9 in ICAM-1 mRNA， IL-6 mRNA and COL-Ⅳ mRNA levels were significantly reduced （P<0.01） and MMP-9 mRNA levels were significantly increased （P<0.05）， ICAM-1， IL-6 and COL-Ⅳ protein expression was significantly reduced （P<0.01） and MMP-9 protein expression was significantly The expression of ICAM-1， IL-6 and COL-Ⅳ proteins was significantly decreased （P<0.01） and MMP-9 protein expression was significantly increased （P<0.01）. ConclusionModified Zhenwutang may regulate the IL-6/MMP-9/COL-Ⅳ signaling pathway， thereby reducing proteinuria， improving renal function， reducing renal pathological damage and delaying the progression of CRF interstitial fibrosis.
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AIM: To investigate the bacterial flora and antibiotic susceptibility testing of conjunctival sac in adolescents after wearing orthokeratology(OK)lens.METHODS:A total of 101 adolescents aged 8 to 14 who admitted to outpatient department of Xi'an No.1 Hospital from September 2021 to August 2022 were recruited in this cross-sectional observational study. There were 51 cases wearing OK Lens(wearing group)and 50 patients not wearing contact lens(non-lens group), the right eye of all patients was selected into the group. The culture of bacterial flora in conjunctival sac between the two groups were compared, the species were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the antibiotic susceptibility testing was carried out for the positive strains cultured in the wearing group.RESULTS:The positive rate of conjunctival sac bacterial cultured in the wearing group and the non-lens group was 68.6%(35/51)and 60.0%(30/50), respectively(P>0.05). In both groups, the bacteria with the highest detection rate were staphylococcus epidermidis and staphylococcus aureus. The sensitivity rates of the strains detected in the wearing group to drugs are as follows: Levofloxacin(98%), Moxifloxacin(98%), Gatifloxacin(98%), Cefuroxime(98%), Cefathiamidine(98%), Rifampicin(98%), Chloramphenicol(96%), Cefoxitin(95%), Clindamycin(80%), Gentamicin(74%), Fusidic acid(72%), Tobramycin(64%), Compound sulfamethoxazole(26%), Mezlocillin(10%), Azithromycin(6%), of which the sensitivity rate of Gram-positive cocci was 100% sensitive to Vancomycin.CONCLUSION:Gram-positive cocci are the main bacteria isolated from conjunctival sac of adolescents after wearing OK Lens. Wearing OK Lens will not significantly increase the positive rate of conjunctival sac bacterial flora. Results of antibiotic susceptibility testing may provide guidance for empirical medication in patients wearing OK lens after eye infection.
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ObjectiveTo study the effect and mechanism of Wuwei Xiaoduyin in treating rat renal mesangial cells （HBZY-1） induced by lipopolysaccharide （LPS） through the nuclear factor-κB （NF-κB） signaling pathway. MethodRat HBZY-1 cells were randomly assigned into the normal group， model group， benazepril （50 μmol·L-1） group， and high- and low-dose （2.75 and 0.69 g·kg-1） Wuwei Xiaoduyin groups. The normal group， model group， and benazepril group were treated with 10% normal rat serum， and the Wuwei Xiaoduyin groups with 10% medicated serum. Except the normal group， the other four groups were treated with LPS （100 ng·mL-1） for modeling in vitro. The changes of cell morphology were observed under optical microscope. The expression of NF-κB p65 was detected by immunofluorescence （IF） method. Methyl thiazolyl tetrazolium （MTT） colorimetry was employed to detect cytotoxicity and cell proliferation. The levels of interleukin-1β （IL-1β）， intercellular adhesion molecule-1 （ICAM-1）， laminin （LN）， and fibronectin （FN） in cell supernatant were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA levels of IL-1β， FN， and NF-κB p65 were measured by real-time fluorescence quantitative PCR. The protein levels of phosphorylated inhibitor of NF-κB kinase β （p-IKKβ）， phosphorylated NF-κB inhibitor （p-IκBα）， and NF-κB p65 were determined by Western blot. ResultCompared with the normal group， the modeling increased cell proliferation （P<0.01）， elevated the levels of IL-1β， ICAM-1， LN， and FN in cell supernatant （P<0.01）， and up-regulated the mRNA levels of IL-1β， FN， and NF-κB p65 （P<0.01） and the protein levels of p-IKKβ， p-IκBα， and NF-κB p65 （P<0.01）. Such changes were recovered by benazepril and Wuwei Xiaoduyin （P<0.05， P<0.01）. ConclusionWuwei Xiaoduyin can mitigate the inflammatory injury of renal mesangial cells induced by LPS by inhibiting the NF-κB signaling pathway.
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AIM: To compare the progress of binocular myopia with orthokeratology lens in different degrees of unilateral myopia in anisometropic children for 1a.METHODS:Retrospective case-control study. The data of 60 unilateral myopic anisometropia children, 8-12-year-old patients who fitted with orthokeratology lens in Xi'an No.1 Hospital from June 2018 to June 2019 were collected. The binocular spherical equivalent(SE)difference ≥2.5D was found in 30 cases in the high anisometropia group, and 1.0D≤ binocular SE difference <2.5D was found in 30 cases of the low anisometropia group. The patients in the two groups were observed and compared after wearing lens for 6mo and 1a of the changes of binocular axial length(AL)and SE without-lens eyes.RESULTS: At the follow-up at 6mo and 1a, the growth of AL in the high anisometropia group with orthokeratology lens was less than that in the low anisometropia group(P<0.001). The amount of growth of AL without-lens eyes and the growth of SE were greater than that in the low anisometropia group(all P<0.001), and the shortening range of the binocular AL in the high anisometropia group was greater than that in the low-degree anisometropia group(P<0.05). The correlation analysis showed that the anisometropia and the shortening of axial difference were correlated(F=0.293, 0.458, all P<0.001).CONCLUSION: In anisometropia children with unilateral myopia, after wearing the orthokeratology lens, the progression in high anisometropia group was slower than low anisometropia after wearing the orthokeratology lens, but the progression of AL without-lens eyes and SE was faster than low anisometropia. Wearing orthokeratology lens can effectively reduce anisometropia, and the greater degree of the anisometropia the more binocular axial difference was narrowed, the more binocular axial difference was narrowed.
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Objective:To study effects of balance and motor function of walking stability training guided by mirror neuron theory for stroke patients with hemiplegia.Methods:A total of 67 stroke with hemiplegia from January 2019 to June 2020 in Heze Municipal Hospital were divided into experimental group (34 cases) and control group (33 cases). All patients were given conventional rehabilitation training, the control group was given body weight support treadmill training, the experimental group was given mirror visual feedback therapy combined with body weight support treadmill training. The training time was 8 weeks. The body balance function, body motor function and life quality were evaluated by Fugl-Meyer Balance scale(FM-B), Berg Balance Scale(BBS), Tinetti Gait Analysis(TGA), Functional Ambulation Category scale(FAC), Stroke-Specific Quality of Life scale (SS-QOL) and compared before and after training between the two groups.Results:There was no significant difference in the score (classification) of FM-B, BBS, TGA, SS-QOL, FAC before training between the two groups( P>0.05). The score (classification) of FM-B, BBS, TGA, FAC after training were (11.35±1.32), (47.33±6.20), (9.32±1.12) points and (3.54±0.61)grades in the experimental group and (9.86±1.25), (42.14±6.35), (8.25±1.32) points and (3.15±0.54) grades in the control group. There were statistically significant of differences( t values were 2.768-4.741, P<0.05). The score of energy, activity ability, emotion, self-care ability, thinking, upper limb function and life quality total score of SS-QOL were (10.31±1.43), (21.46±4.45), (17.64±3.36), (17.32±3.15), (16.23±2.21), (159.11±16.34) points in the experimental group and (8.46±1.21), (17.32±3.24), (15.35±3.11), (14.78±2.32), (8.45±1.25), (13.45±1.54), (141.65±17.24) points in the control group. There were statistically significant of differences( t values were 2.893-7.096, P<0.05). Conclusions:Walking stability training based on mirror neuron theory help to restore limb balance and motor function, and improve life quality in stroke patients with Hemiplegia.
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Patients with oncogenic driver alterations of non-small cell lung cancer (NSCLC) can benefit from targeted therapy, but acquired resistance is inevitable ultimately. Epigenetic modifications, including DNA methylation, histone modifications, non-coding RNA-mediated regulate and chromatin remodeling, are important mechanisms of acquired resistance in targeted therapy of NSCLC. In recent years, studies have found that epigenetic modifications can effectively reverse drug resistance. Targeted therapy combined with epigenetic modifications may become a promising therapeutic strategy. Here, we review the progress of epigenetic mechanism in acquired resistance of targeted therapy in NSCLC, hoping to provide ideas for screening dominant population and overcoming resistance. .
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Humans , Carcinoma, Non-Small-Cell Lung/genetics , DNA Methylation , Epigenesis, Genetic , Lung Neoplasms/geneticsABSTRACT
Objective@#To investigate the diagnostic value of ultrasonography.in elderly patients with gastric tumors.@*Methods@#A total of 42 elderly patients with gastric tumors confirmed by surgery pathology underwent oral contrast-enhanced ultrasonography and electronic gastroscopy.The diagnostic effect of oral contrast-enhanced ultrasonography and electronic gastroscopy were compared based on the gold standard of surgical pathology.@*Results@#Of the 42 elderly patients with gastric tumors, 40 patients were diagnosed by oral contrast-enhanced ultrasonography and the surgical pathology(40/42, 95.2%), with Kappa value of 0.812.And 38 cases were diagnosed by electronic gastroscopy and the surgical pathology(38/42, 90.5%), with Kappa value of 0.718(P<0.05). Based on gold standard of surgical pathology results, the accuracies of T1, T2, T3 and T4 stage by oral contrast-enhanced ultrasonography were 95.0%(38/40), 95.0%(38/40), 100.0%(40/40)and 100.0%(40/40), respectively.The sensitivity and specificity of T3 and T4 stage for oral contrast-enhanced ultrasonography reached to 100.0%.The diagnostic accuracies of N0, N1-N3, M0 and M1 by oral contrast-enhanced ultrasonography were 95.0%(38/40), 95.0%(38/40), 100.0%(40/40)and 100.0%(40/40).@*Conclusions@#Oral contrast-enhanced ultrasonography is a simple, painless and non-invasive method for the diagnosis of gastric tumors in the elderly.And its qualitative coincidence rate of the tumors diagnosis is similar to that of gastroscopy.So it can be used as the first choice for the examination of gastric tumors in the elderly.Oral contrast-enhanced ultrasonography has a high diagnostic accuracy on TNM staging of gastric tumor.When combined with electronic gastroscopy, it may provide a more reliable basis for the choice of evaluation of treatment and prognosis in elderly patients with gastric tumor.
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OBJECTIVE@#Using the CT three-dimensional reconstruction to measure the activity degree of atlanto-occipital joint and the atlantoaxial joint in different directions and its coupling movement in healthy volunteers, and three dimensional motion range of the maximum rotation position of the upper cervical spine of cervical spondylosis patients, and to analyze the differences, verifing the reliability of the method at the meantime.@*METHODS@#From January 2014 to June 2015, 20 healthy adult subjects(healthy adult group), and 26 patients with cervieal spondylosis(cervical spondylosis group) were selected. In healthy adult group, there were 11 males and 9 females, aged from 22 to 26 years old with an average of (24.0±1.2) years, and in cervical spondylosis group, there were 24 males and 2 females, aged from 36 to 72 years old with an average of (52.8±8.6) years. Healthy adults underwent CT examination in neutral position, maximum right rotation, maximum right lateral bending, maximum flexion and extention, and cervical spondylosis patients underwent CT examination in neutral position, maximum right rotation. Then the software Mimics was used to reconstruct occiput (Oc), atlas(C1) and axial(C2) vertebral three-dimensional image. Three virtual non-collinear markers were positioned on prominent structures of foramen magnum, C1 and C2. The 3D spatial coordinates of these virtual anatomical markers entail the definition of an anatomical local coordinate system which represent the position and orientation of the bones. Segmental motions were calculated using Eulerian angle in three major planes, and the difference between cervical spondylosis group and healthy adult group were compared. Due to the inaccuracy in anatomical landmark idenrification, two groups were measured 3 times, and the reliability of the experimental metnod was verified by the intra-group correlation (intra-group ICC) and the inter-group correlation coefficient(inter-group ICC).@*RESULTS@#Reliability verification results:the intra-group ICC and inter-group ICC results were all above 0.90, and the measurement method had high reliability. Three-dimensional activity of the upper cervical spine in healthy adults:the atlanto-occipital joint had(-6.8±1.5)° coupled left lateral bending and (8.9±2.0)° coupled extension in the maximum right rotation position, and the motion of atlanto-occipital joint had low activity[maximum was(5.3±2.6)°] in the remaining 3 positions; the rotation of atlanto-axial joint was(37.9±5.1)°, accounting for 52.34% of the total cervical spine activity[(72.4±5.0)°] in the maximum right rotation position, and rotational motion was still prominent in the remaining three positions. The relative translations of the upper cervical spine in all direction were small. The average axial rotation angle [(62.0±3.4)] ° of the total cervical spine in cervical spondylosis group was significantly lower than that in the healthy adult group, but the mean axial rotation angles of the atlanto-occipital and the atlantoaxial joint were not significantly different from those of the healthy adults(>0.05).@*CONCLUSIONS@#The three-dimensional CT reconstruction method has high reliability, which can be applied to measure the movement of spine. The upper cervical spine contributed the most to the direction of rotation, and the movement in all directions are accompanied by coupled motion in the other direction. There was no significant difference in the rotation of the upper cervical spine between cervical spondylosis patients and normal subjects.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Atlanto-Axial Joint , Biomechanical Phenomena , Cervical Vertebrae , Imaging, Three-Dimensional , Range of Motion, Articular , Reproducibility of Results , Rotation , Spine , Tomography, X-Ray ComputedABSTRACT
Objective To investigate the diagnostic value of ultrasonography.in elderly patients with gastric tumors.Methods A total of 42 elderly patients with gastric tumors confirmed by surgery pathology underwent oral contrast-enhanced ultrasonography and electronic gastroscopy.The diagnostic effect of oral contrast-enhanced ultrasonography and electronic gastroscopy were compared based on the gold standard of surgical pathology.Results Of the 42 elderly patients with gastric tumors,40 patients were diagnosed by oral contrast-enhanced ultrasonography and the surgical pathology(40/42,95.2％),with Kappa value of 0.812.And 38 cases were diagnosed by electronic gastroscopy and the surgical pathology(38/42,90.5 ％),with Kappa value of 0.718 (P ＜ 0.05).Based on gold standard of surgical pathology results,the accuracies of T1,T2,T3 and T4 stage by oral contrast-enhanced ultrasonography were 95.0％ (38/40),95.0％ (38/40),100.0％ (40/40) and 100.0％(40/40),respectively.The sensitivity and specificity of T3 and T4 stage for oral contrastenhanced ultrasonography reached to 100.0 ％.The diagnostic accuracies of N0,N1-N3,M0 and M1 by oral contrast-enhanced ultrasonography were 95.0 ％ (38/40),95.0 ％ (38/40),100.0 ％ (40/40) and 100.0％(40/40).Conclusions Oral contrast-enhanced ultrasonography is a simple,painless and non-invasive method for the diagnosis of gastric tumors in the elderly.And its qualitative coincidence rate of the tumors diagnosis is similar to that of gastroscopy.So it can be used as the first choice for the examination of gastric tumors in the elderly.Oral contrast-enhanced ultrasonography has a high diagnostic accuracy on TNM staging of gastric tumor.When combined with electronic gastroscopy,it may provide a more reliable basis for the choice of evaluation of treatment and prognosis in elderly patients with gastric tumor.
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Objective To study and compare the efficacy of surgery and conservative treatments for patients with critical(30 to 50 ml) spontaneous deep supratentorial intracerebral hemorrhage. Methods The clinical data of 420 patients diagnosed as spontaneous deep supratentorial intracerebral hemorrhage in Affiliated Hospital of Yan'an and Central Hospital of Baoji from August 2011 to September 2016 in this study were retrospectively analyzed. All patients were divided into surgery group(210 cases) and conservative group(210 cases) according to the difference of treatment methods. Large trauma craniotomy was used by the operation group, external ventricular drainage as the standby in necessary. The treatment of conservative group mainly included reduction of intracranial pressure, blood pressure management, prevention of complications and other individualized treatment. The primary outcome was neurological functional status of patients at 6 months of discharging, and the secondary outcomes included the mortality at 1 month and 6 months of discharging, and complications. Continuous data were presented as (x)± s and were compared using the t test, while categorical data were presented as number and percentage and were compared using the χ2 test. Results No obvious difference on mRS score and NIHSS score occurred between two groups(All P>0.05). Stratified analysis, patients with midline shift >5 mm and with intraventricular hemorrhage were more likely to have a favorable outcome from surgery compared with conservative group(18.2％ vs.8.1％, X =4.099, P=0.043; 17.3％ vs.5.0％, χ2 =4.836, P=0.028). The mortality at 1 month days and 6 months in surgery group were significantly lower than that in conservative group(19.0％ vs.31.0％, X2 =7.937, P=0.005; 23.8％ vs.36.7％, X2=8.228, P=0.004). Meanwhile, the incidence of pulmonary infection and with long-term pipe in surgery group were significantly higher than those in conservative group(31.9％ vs.15.7％, X2 =15. 173, P=0.000; 28.6％ rs.19.5％ =4.706, P=0.030). Conclusion Surgery can reduce the 1 month and 6 months mortality in patients with spontaneous deep supratentorial hemorrhage and improve the functional outcomes in patients with large midline shift or with intraventricular hemorrhage compared with conservative treatment.
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Objective@#To observe the uptake of 99Tcm-3 polyethylene glycol Arg-Gly-Asp dimer (3PRGD2) in rat models of rheumatoid arthritis (RA), in order to provide theoretical foundation for early diagnosis of RA.@*Methods@#The healthy female SD rats were divided into collagen induced arthritis (CIA) group (n=100), osteoarthritis (OA) group (n=20) and control group (n=20). Bovine collagen type Ⅱ emulsion was used for arthritis induction to establish CIA models. OA models were established by injection of L-cysteine and papain. Gamma imaging was performed before and 15 d, 30 d after the model establishment. The mediastinum was selected as non-target (NT) area and the target (T)/NT ratios were calculated. The serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-α (TNF-α), αVβ3 before and after the model establishment were measured. Pathological and immunohistochemical detection were performed. One-way analysis of variance, Pearson and Spearman correlation analyses were used to analyze the data.@*Results@#There were 58 CIA models successfully established. Before the model establishment, T/NT ratios of CIA group, OA group and control group were 0.215±0.049, 0.210±0.050, 0.209±0.051, respectively (F=0.093, P>0.05). The T/NT ratios of the above three groups were 0.405±0.230, 0.223±0.045, 0.211±0.049 (F=12.601, P<0.05) 15 d post-model establishment, and those were 0.572±0.182, 0.238±0.045, 0.212±0.055 (F=65.147, P<0.05) 30 d post-model establishment. T/NT ratios of CIA group were positively correlated with the levels of serum VEGF, TNF-α, αVβ3, pathological score and the levels of immunohistochemical markers (VEGF, TNF-α, αVβ3, CD31, CD34; r values: 0.391-0.721, rs values: 0.365-0.669, all P<0.05).@*Conclusion@#99Tcm-3PRGD2 can be specifically up-taken by RA lesions in rat models, thus has the potential in early synovial neovascularization imaging of RA.
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Objective To investigate the effects of α-linolenic acid (ALA) on acute inflammation and neurological functional recovery of mice with traumatic brain injury (TBI). Methods The C57BL6/N mice were divided into high ALA dietary group (20 pregnant and 88 new-born mice) and low ALA dietary group (20 pregnant and 84 new-born mice) in this study. The contents of polyunsaturated fatty acids in the brain of the two groups were detected by gas chromatography. Mouse TBI model was established by control cortical impact method. The expression levels of inflammatory cytokines and cellular markers of the two groups were detected by reverse transcription-quantitative polymerase chain reaction and enzyme linked immunosorbent assay or Western Blotting at 0, 4, 24 and 96 h after TBI, respectively. The neurological functions were analyzed by the rotarod, beam walk test and fear conditioning experiment. Results The content of brain docosahexoenoic acid (DHA) was significantly higher in the high ALA dietary group than in the low ALA dietary group (15.48±1.20% vs 9.98±1.10%, P<0.05). The expression levels of TNF-α, IL-1β , IL-6 and CCL12 were lower in high ALA groups than in low ALA group after TBI (P<0.05). The motor function recovery 24 h after TBI was faster in the high ALA dietary group than in the lower ALA diet group. The cognitive function 24 h after TBI was better in the high ALA dietary group than in the low ALA group. Conclusion Increasing DHA levels in the brain can reduce acute inflammation and improve neurological function recovery after TBI.
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This study was aimed to investigate the effects of decitabine (DAC) on proliferation and apoptosis of leukemia NB4 and K562 cells. The proliferation inhibition of DAC on NB4 and K562 cells was detected by Trypan blue staining. After treatment of DAC at different concentrations, the changes of cell cycle and CD11b expression was determined by flow cytometry. The cell morphological changes were observed by Wright's staining. The DNA ladder was used to detect cell apoptosis. The results indicated that DAC significantly inhibited the proliferation of NB4 and K562 cells in dose-and time-dependent manner. The median inhibitory concentration (IC50) of DAC-treated NB4 and K562 cells for 72 h was 0.113 µmol/L and 0.138 µmol/L, respectively. After treating these two cell lines with DAC at different concentration for 72 h, the cell ratio in G0/G1 phase significantly increased, while the cell ratio in S phase obviously decreased in 0.15 µmol/L DAC group (P < 0.05). The expression levels of myeloid differentiation antigen CD11b of both cell lines significantly increased in contrast to the control group (P < 0.05). The cell morphology detected by Wright's staining displayed partial differentiation and apoptosis after treating NB4 and K562 cells with DAC for 48 h. Typical apoptotic DNA ladder was observed in 0.15 µmol/L DAC group at 48 h. It is concluded that DAC can inhibit NB4 and K562 cell proliferation, induce cell differentiation and apoptosis, but more obviously for NB4 cells.
Subject(s)
Humans , Apoptosis , Azacitidine , Pharmacology , Cell Differentiation , Cell Line, Tumor , Cell Proliferation , K562 CellsABSTRACT
<p><b>OBJECTIVE</b>To study the demethylation effect of arsenic trioxide (As2O3) on ERα-negative human breast cancer MDA-MB-435s cells and its possible mechanisms, and to observe its treatment efficacy in combination with tamoxifen (TAM) after ERα re-expression.</p><p><b>METHODS</b>MTT assay was used to examine the inhibitory effect of As2O3 treatment alone or in combination with TAM on cell proliferation. A nude mouse xenograft model was used to further examine the treatment efficacy in vivo. MSP was used to detect the methylation status of ERα gene after treated with As2O3 in MDA-MB-435s cells and the transplanted tumor tissues. RT-PCR was used to detect the mRNA expression of DNMT1 and Erα. Western bolt was used to detect the DNMT1 and ERα protein expression. The diameter of xenograft tumors was measured weekly, and the tumor growth curve was drawn.</p><p><b>RESULTS</b>The level of proliferation of the MDA-MB-435s cells was significantly suppressed after treatment with different concentration of As2O3 alone or As2O3 combined with TAM, and the 4 µmol/L As2O3 + TAM treatment for 72 h showed the highest inhibition rate (62.6%). 1, 2, 4 µmol/L As2O3 had demethylation effect on MDA-MB-435s cells, and the DNMT1 mRNA and protein expression was inhibited and accompanied by ERα mRNA and protein re-expression. The unmethylation specific bands of ERα gene were enhanced after treated by As2O3 alone or As2O3 combined with TAM in the xenograft tumors. The expression of DNMT1 mRNA and protein was inhibited, and accompanied by ERα mRNA and protein re-expression. An significant decrease of volume and weight of the xenograft tumors in the As2O3 treated alone or combined with TAM groups was observed compared with those of the normal saline group or TAM alone group (P < 0.05), and the 4 mg/kg As2O3 + TAM group had the highest inhibition rate of tumor weight (79.5%) and volume (76.4%).</p><p><b>CONCLUSIONS</b>ERα can be re-expressed in ERα-negative breast cancer MDA-MB-435s cells after treated with As2O3 by inhibiting the DNMT1 activity. MDA-MB-435s cells are re-sensitized to endocrine therapy after ERα re-expression. As2O3 combined with TAM may provide a new therapeutic approach for patients with ERα-negative breast cancer in the clinic.</p>
Subject(s)
Animals , Female , Humans , Mice , Antineoplastic Agents , Pharmacology , Antineoplastic Agents, Hormonal , Antineoplastic Combined Chemotherapy Protocols , Pharmacology , Arsenicals , Pharmacology , Breast Neoplasms , Genetics , Metabolism , Pathology , Cell Line, Tumor , Cell Proliferation , DNA (Cytosine-5-)-Methyltransferase 1 , DNA (Cytosine-5-)-Methyltransferases , DNA Methylation , Dose-Response Relationship, Drug , Estrogen Receptor alpha , Genetics , Metabolism , Mice, Inbred BALB C , Mice, Nude , Oxides , Pharmacology , RNA, Messenger , Metabolism , Tamoxifen , Tumor Burden , Xenograft Model Antitumor AssaysABSTRACT
The aim of this study was to observe the electrophysiological characteristics of the isolated rat muscle spindle. The muscle spindle was isolated from rat soleus and the afferent discharge of the isolated muscle spindle was recorded by air-gap technique. In the basic physiological salt solution, the spontaneous impulses of muscle spindle were at a lower level with irregular intervals. The mean frequency of afferents was (51.78 ± 25.63) impulses/1 000 s (n = 13). The muscle spindle afferents were significantly increased and maintained over time by the addition of certain amino acids during the observation. The number of the action potential recorded per 1 000 s was 200-1 000 [mean: (687.62 ± 312.56) impulses/1 000 s, n = 17]. In addition to the typical propagated action potential, a large number of abortive spikes were observed. The results indicate that the activities of isolated muscle spindles in rats can be well maintained by the addition of certain amino acids. The results initially establish and provide the possibility for further research conducted in isolated rat muscle spindles.
Subject(s)
Animals , Female , Rats , Action Potentials , Physiology , Amino Acids , Pharmacology , Electrophysiological Phenomena , In Vitro Techniques , Muscle Spindles , Physiology , Muscle, Skeletal , Physiology , Rats, Sprague-Dawley , Sodium Chloride , PharmacologyABSTRACT
The present study aimed to study the changes of neurotrophin-3 (NT-3) expression of intrafusal muscle fibers in rat soleus muscles under simulated weightlessness. The tail-suspension (SUS) rat model was used to simulate weightlessness. Forty mature female Sprague-Dawley rats were randomly assigned to ambulatory control (CON), 3-day SUS, 7-day SUS, 14-day SUS and 21-day SUS groups. Immunohistochemistry ABC staining method and enzyme linked immunosorbent assay (ELISA) were used to detect the NT-3 expression of intrafusal muscle fibers in rat soleus muscles. The results from the immunohistochemistry staining technique showed that the extrafusal muscle fibers did not exhibit the NT-3-like immunoreactivity, and NT-3-like immunoreactivity was mainly expressed in nuclear bag 1 and nuclear bag 2 fibers of the muscle spindles. The ELISA results showed that the expression quantity of NT-3 in rat soleus muscles in control, 3-day SUS, 7-day SUS, 14-day SUS and 21-day SUS groups were (14.23±1.65), (14.11±1.53), (13.09±1.47), (12.45±1.51) and (9.85±1.52) pg/mg of tissue respectively. Compared to the control group, the expression quantity of NT-3 was significantly decreased after 14 days of SUS (P<0.05). After 21 days of SUS, the NT-3 expression was further reduced (P<0.01). These results suggest that simulated weightlessness induces an obvious decrease in the NT-3 expression level of intrafusal fibers in rat soleus muscles. Accompanying the simulated weightlessness extension, NT-3 expression in rat soleus muscle spindles is progressively decreased. These changes may contribute to the proprioceptive adaptations to microgravity.
Subject(s)
Animals , Female , Rats , Down-Regulation , Hindlimb Suspension , Muscle Spindles , Metabolism , Muscle, Skeletal , Metabolism , Neurotrophin 3 , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Weightlessness SimulationABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of 100 Hz sinusoidal vibration on H reflex and M wave in rat soleus muscle following immobilization.</p><p><b>METHODS</b>The immobilization of rat soleus muscle was induced as a disuse muscle model, and 100 Hz sinusoidal vibration was generated by a vibrator and applied to the immobilized soleus muscle, then the changes of H reflex and M wave in muscle were observed after 14 d.</p><p><b>RESULTS</b>Compared to control, after 14 d of immobilization M(max) in soleus muscle decreased (P<0.01), stimulus threshold and S(max) increased (P<0.01); Hmax and H(max)/M(max) decreased (P<0.05, S(max) increased (P<0.05). Compared to immobilized soleus muscle, after 14 d of immobilization with 100 Hz sinusoidal vibration, the M(max) increased(P<0.01), stimulus threshold and S(Mmax) decreased (P<0.05), H(max) (P<0.01) increased and H(max)/M(max) increased (P<0.05).</p><p><b>CONCLUSION</b>100 Hz sinusoidal vibration plays a significant antagonist role against the changes in H reflex and M wave in rat soleus muscle following immobilization.</p>
Subject(s)
Animals , Female , Rats , Electromyography , H-Reflex , Physiology , Hindlimb Suspension , Muscle Contraction , Physiology , Muscle Spindles , Physiology , Muscle, Skeletal , Physiology , Rats, Sprague-Dawley , VibrationABSTRACT
<p><b>OBJECTIVE</b>To analyze the differential proteomics in human umbilical cord mesenchymal stem cells (MSC) induced by chemical hypoxia-mimetic agent cobalt chloride (CoCl(2)) by two-dimensional gel electrophoresis (2-DE) and mass-spectrometry.</p><p><b>METHODS</b>2-DE was performed to separate proteins from treated and untreated human umbilical cord MSC with CoCl(2). 2-DE images were analyzed by ImageMaster 2D Platinum software 6.0. The differential expressed proteins was identified by MALDI-TOF-MS. The differential proteins were classified based on their functions.</p><p><b>RESULTS</b>2-DE reference patterns of CoCl(2) treated human umbilical cord MSC were established. A total of twenty-six differential proteins were identified, of them eleven proteins were up-regulated and fifteen down-regulated. Their biological functions involved in carbohydrate metabolism, protein metabolism and modification, lipid metabolism, coenzyme and prosthetic group metabolism, cell cycle, immunity and defense, cell structure and motility, signal transduction, protein targeting and localization, neuronal activities, muscle contraction, etc. Peroxiredoxin1 (Prdx) was down-regulated, whereas alpha-enolase (ENO1) and vesicle amine transport protein 1 homolog (VAT1) up-regulated.</p><p><b>CONCLUSION</b>The effects of hypoxia on human umbilical cord MSC were participated by multiple proteins and involved in multiple functional pathways.</p>
Subject(s)
Humans , Cobalt , Pharmacology , Mesenchymal Stem Cells , Metabolism , Proteome , Proteomics , Umbilical Cord , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the mechanism of apoptosis of EC9706 tumor-bearing nude mice induced by all-trans retinoic acid (ATRA).</p><p><b>METHODS</b>Human esophageal carcinoma cell line EC9706 cells were inoculated into nude mice to establish the solid tumor model. The tumor models were divided into the following groups: ATRA group, fluorouracil group, the two-drugs combination group, and with an equal volume fraction of solvent as the control group. The nude mice were sacrificed after 10 days of medication. TUNEL staining was used to detect cell apoptosis. RT-PCR was used to detect the expression level of mRNA and immunohistochemistry was used to detect the expression level of protein of caspase-3 and survivin, the apoptosis-related genes in the tumor tissue.</p><p><b>RESULTS</b>The apoptosis rates of the ATRA group, 5-Fu group and ATRA + 5-Fu group were 44.3%, 39.7% and 91.0%, respectively. There was a significant difference in comparison with the control group (0.7%), and the ATRA group had no significant difference compared with that of the fluorouracil group (P > 0.05), but the apoptosis rate of the two-drugs combination group was significantly higher than that in the two single-drug groups (P < 0.05). The average gray value of caspase-3 protein expressed in the control group was 46.12 ± 0.33 and the relative expression of caspase-3 mRNA was 0.14 ± 0.03, both were significantly lower than that in the ATRA group, 5-Fu group and the two-drugs combination group (P < 0.05). The average gray value of survivin protein expressed in the control group was 96.07 ± 0.13 and the relative expression of survivin mRNA was 0.84 ± 0.04, both were significantly higher than those of other groups (P < 0.05). The ATRA group had no significant difference compared with the fluorouracil group (P > 0.05), but the two-drugs combination group was significantly different compared with the single-drug groups (P < 0.05).</p><p><b>CONCLUSION</b>Apoptosis in the EC9706 tumor cells in nude mice can be induced by ATRA. The mechanism may be related with down-regulation of the level of survivin gene expression and up-regulation of the level of caspase-3 gene expression.</p>
Subject(s)
Animals , Female , Humans , Male , Mice , Antimetabolites, Antineoplastic , Pharmacology , Antineoplastic Agents , Pharmacology , Apoptosis , Caspase 3 , Genetics , Metabolism , Cell Line, Tumor , Esophageal Neoplasms , Metabolism , Pathology , Fluorouracil , Pharmacology , Inhibitor of Apoptosis Proteins , Genetics , Metabolism , Mice, Nude , Neoplasm Transplantation , RNA, Messenger , Metabolism , Tretinoin , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the protective action of Epimedium against chemotherapy-induced damage to rat epididymides.</p><p><b>METHODS</b>Fifty 60-day-old male rats were divided into a control, a model and a treatment group. Procarbazine was injected into the abdominal cavity of the model rats at the dose of 30 mg/(kg x d). In addition to procarbazine, Epimedium was given intragastrically to the treatment group. The changes in the ultrastructure of the epididymis were observed after 10 and 20 days.</p><p><b>RESULTS</b>Electron microscopy showed that the chemotherapy-induced damages to the epididymal epithelia mainly included cell swelling, local cavitation of mitochondria, tumor-like change in nucleoli, agglutination of marginal translocation of heterochromatin and cell apoptosis. The damage to the epithelial ultrastructure was slight in the treatment group as compared with the model rats. Chemotherapy significantly affected sperm concentration, sperm viability and sialic acid (SA), which were (15.59 +/- 4.01) x 10(6)/ml, (76.71 +/- 10.11)% and (19.38 +/- 9.34) g/mg prot in the model group in comparison with (10.63 +/- 3.82) x 10(6)/ml (P < 0.01), (60.03 +/- 7.54)% (P < 0.01) and (13.62 +/- 7.81) g/g prot (P < 0.05) in the control. Epimedium significantly increased sperm viability in the treatment group (60.03 +/- 7.54)% as compared with the model rats (69.90 +/- 12.58)% (P < 0.05).</p><p><b>CONCLUSION</b>Epimedium can lessen chemotherapy-induced damage to the epididymis and protect the reproductive function of rats.</p>