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Four new diphenyl ethers, named epicoccethers K-N (1-4), were purified from the fermentation medium of a fungus Epicoccum sorghinum derived from Myoporum bontioides, and identified through HR-ESI-MS and NMR spectral analysis. Except that compound 1 showed moderate antifungal activity against Penicillium italicum and Fusarium graminearum, the other three compounds showed stronger activity against them than triadimefon. All of them showed moderate or weak antibacterial activity towards Staphylococcus aureus and Escherichia coli with O6 and O78 serotypes except that 3 was inactive to E. coli O6.
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Anti-Bacterial Agents/pharmacology , Antifungal Agents/chemistry , Ascomycota , Escherichia coli , Microbial Sensitivity Tests , Molecular Structure , Phenyl Ethers/chemistryABSTRACT
When this paper was first published the following ethical statement was omitted in error: Animal experiments were conducted in accordance with the guidelines of Laboratory Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (TCM-LAEC2019071). The authors would like to apologise for any inconvenience caused. DOI of original article: https://doi.org/10.1016/j.chmed.2018.10.002
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Objective:To evaluate the value of a novel multiphase three-dimensional deep learning neural network of computer-aided diagnosis (CAD) used in LDCT lung cancer screening.Methods:Eight thousand eight hundred and fifty volunteers with 1 111 nodules were enrolled in the lung cancer screening from November of 2013 to December of 2017, and the baseline LDCT imaging data of volunteers accompanied with clinical information were retrospectively analyzed. All volunteers in this study were designed to receive LDCT test at least once. All the imaging of volunteers were read through the methods of visual detectioin (VD), CAD, and VD Combined CAD. The criteria of the true pulmonary nodule was determinated by the consistent opinion of two specialists in chest imaging（in case of disagreement, the decision should made by the third chief physician). In terms of the numbers, types or Lung-RADS categories of nodules, the detection rate, missed diagnosis rate and false positive rate of pulmonary nodules or lung cancer among three methods were compared, and the rates between groups were compared by χ 2test. Results:Compared with VD or CAD ,the detection rate of nodules in the CAD combined VD was significantly increased (95.7% , 94.2%, vs. 80.1% P<0.05 ), and the rate of missed diagnosis was significantly reduced (5.8%, 4.3% vs. 19.9% ，χ2=101.650, 128.500 ，P<0.05); Compared with VD, the methods of CAD or VD combined CAD significantly increased the the detection rates of Lung-RADS categories (χ2 =25.083,23.449, P=0.000, 0.000) or different types of nodules (χ2=6.955,6.821, P=0.031, 0.033), but there was no statistically significant difference between CAD and VD combined CAD for Lung-RADS categories and different types of nodules (all P>0.05); Compared with VD and VD combined CAD, the positive prediction rate of CAD for lung cancer was significantly reduced, and the rate of missed diagnosis and false positive rate were significantly increased, but there was no significant difference between VD and VD combined CAD in the prediction rate, missed diagnosis rate and false positive rate of lung cancer. Conclusion:The method of CAD combined VD can reduce the detection of false positive nodules and improve the detection rate of true pulmonary nodules,which is the preferred method using in LDCT lung cancer screening for city population.
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Objective@#To analyze the teaching environment of primary, middle and high schools in Suzhou, and to explore the relevance of students’ myopia with teaching environment, as well as the key problems in teaching environment.@*Methods@#1 253 classrooms in 204 schools rom 10 districts and counties in Suzhou were selected by proportionate stratified sampling. The qualified rate of each indices was described and the teaching environment by county, learning stage were compared by using chi-squared test. The Spearman test is used to explore correlations between myopia with monitoring indicators.@*Results@#The qualified rate of the distance from lamp to table was the highest (98.56%), while matching rate of student desk and chair was the lowest(0.96%). Minimum lamp table spacing, blackboard average illumination, reflectance from blackboard wall, classroom per capita area, classroom microclimate differed significantly in urban and rural (P<0.05); daylighting coefficient, reflectance from walls, average illumination of desktop and blackboard, classroom per capita area, and microclimate indicator were statistically different by learning stages(r=0.42,P<0.05). Prevalence of myopia correlated with the index of lighting.@*Conclusion@#Qualified desks and matched chairs, as well as the daylighting design in classroom should receive more attention in Suzhou. The findings lend support to identification of key health priorities and formulation of the localized health practice for health administrations.
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Objective: Monitoring the unbound drug concentration in blood in combination therapy is necessary, because its concentration determines the efficacy of drug therapy. This study was designed to explore the effect of Dan Hong Injection (DHI) on the unbound warfarin using two approaches including an in vivo pharmacokinetic and in vitro fluorescence studies. Methods: The effect of DHI on the pharmacokinetic properties of the unbound warfarin was investigated by a microdialysis sampling method coupled with LC–MS/MS. The effect of DHI and salvianolic acid B (SaB) on warfarin binding with bovine serum albumin (BSA) was conducted by fluorescence spectrometry. Results: The AUC0-tn of warfarin with DHI group was higher than that of warfarin alone group. The result showed that DHI could increase the concentration of unbound warfarin in rat blood, which may be due to the competition between warfarin and DHI as well as its components binding to serum albumin. The competition process was demonstrated by fluorescence study. Conclusion: Combination therapy of DHI with warfarin could enhance the release profile of warfarin from serum protein.
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Anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) is usually a multisystem disorder,and pulmonary renal syndrome is a common presentation.Patients with AAV are less likely to experience relapse when they progress to end-stage renal disease (ESRD).We report a rare case of diffuse alveolar hemorrhage (DAH) in relapsing AAV after eight years of haemodialysis.A 58-year-old woman was admitted to our hospital with the chief complaints of dyspnea and hemoptysis accompanied by anemia,fever,fatigue,and weight loss.She had elevated anti-myeloperoxidase (MPO) titer.The computer tomoghraphy showed diffuse alveolar hemorrhage.After the recurrent episode of AAV was diagnosed,she underwent the following therapy:Plasmapheresis was initiated within 24 h after admission,3 000 mL of plasma was removed per session,and the anticoagulation of citrate was applied during plasmapheresis.Five plasmapheresis treatments were performed,and after three apheresis sessions,the pulmonary hemorrhage ceased.Other treatments included a methylprednisolone bolus,tapered to oral prednisone and cyclophosphamide.Regular hemodialysis was scheduled.These treatments resulted in resolution of the inflammatory symptoms,DAH improved.Her anti-MPO level decreased.The patient was discharged in good condition.AAV with DAH is usually acute at the onset and is generally a condition with high morbidity and substantial mortality.Therefore,prompt diagnosis and aggressive treatment are needed to improve survival.
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Objective To investigate the effect of rosiglitazone on the proliferation and apoptosis of colon cancer cells and the changes in the AKT/GSK3β signaling pathway. Methods Different concentrations of rosiglitazone ( 20. 0μmol/L, 40. 0 μmol/L, 80. 0 μmol/L) were used to treat colon cancer HT29 cells and HCT116 cells. Cell proliferation was detected by MTT assay. Annexin V FITC/PI cell death detection kit was used to test the cell apoptosis rate. The expression of apoptotic protein Bcl-2, Bax and Akt, GSK3β were detected by Western blot. Results Different concentrations of rosiglitazone had different effect on the proliferation of colon cancer cells compared with the blank control group, and showed a dose dependence (P< 0. 01). With the increase of rosiglitazone dose, the apoptosis-inducing effect @was increased dose-dependently (P< 0. 01). When the cells were treated with rosiglitazone for 48 h, the expressions of Bcl-2/Bax, p-GSK3β, and p-Akt were significantly decreased compared with the blank control group (P< 0. 01), but the expression level of Akt and GSK3β was not significantly different compared with the blank control group ( P > 0. 05 ) . Conclusions Rosiglitazone significantly induces apoptosis and inhibits the proliferation of HT29 cells. It may be via inhibiting Akt/GSK3β signaling pathway and change the ratio of Bcl-2/Bax.
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Objective To observe the clinical efficacy of electroacupuncture (EA) in treating knee osteoarthritis (KOA).Method Sixty KOA patients were randomized into a treatment group and a control group by using random number table, 30 cases each. The control group was intervened by oral administration of Celecoxib capsules, while the treatment group was given EA, 14 d as a treatment course. The changes of relevant cytokines [apelin, tumor necrosis factor (TNF)-α, TNF soluble receptor (TNFsR)-Ⅰ, TNFsR-Ⅱ, interleukin (IL)-1β, and IL-6] in serum of the two groups were observed.Result The intra-group comparisons of the total score, and the scores of pain, stiffness and dysfunction of the Western Ontario and McMaster Universities (WOMAC) osteoarthritis index showed significant differences in both groups (P<0.05); there were significant between-group differences in comparing the total score, and the scores of pain, stiffness and dysfunction of the WOMAC index after the treatment (P<0.05). The Visual Analogue Scale (VAS) scores were changed significantly after the intervention in both groups (P<0.05); there was nosignificant difference in comparing the VAS score between the two groups after the treatment (P>0.05). The levels of IL-6, TNF-α and TNFsR-Ⅰ were significantly changed after the treatment in both groups (P<0.05); the level of IL-1β was markedly changed after the intervention in the control group (P<0.05); there was a significant change in the level of apelin after the intervention in the treatment group (P<0.05), and there was a significant difference in comparing the level of apelin between the two groups after the treatment (P<0.05).Conclusion EA can produce a satisfactory efficacy in treating KOA; it can significantly improve the symptoms and signs, and mitigate pain and symptoms through regulating the expressions of cytokines.
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Objective To screen the differentially expressed microRNAs (miRNAs) in colorectal cancer tissues with high expression of gastrin (GAS) by gene chip technique. Methods The level of GAS in tumor tissues from 71 patients with colorectal cancer was detected by enzyme linked immumosorbent assay (ELISA), and the relationship between positive expression of GAS and clinicopathological features was analyzed. Human colorectal cancer tissues with high positive GAS expression (high-expression group, n=4) or negative GAS expression (control group, n=4) were analyzed by miRNA microarray, and the selected differentially expressed miRNAs were validated by qPCR. Results There were 17 cases (23.9%) with GAS content ≥50.00 pg/g (positive GAS expression), while the remaining 54 cases (76.1%) had negative GAS expression. The expression level of GAS was related to the degree of differentiation, Dukes stage and histological type (P200.00 pg/g) compared with the control group, of which 159 miRNAs were up-regulated and 77 were down-regulated. A total of 24 miRNAs were screened out, which differentially expressed more than 3 folds in the high-expression group, with 17 up-regulated and 7 down-regulated. Three up-regulated or down-regulated miRNAs were selected for qPCR verification, and the results were consistent with the microarray analysis. Conclusion The miRNAs are differentially expressed in the colorectal cancer tissues with high expression of GAS, and they would be potential targets for the treatment of the colorectal cancer with high GAS expression.
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Objective To observe the clinical efficacy of electroacupuncture (EA) in treating knee osteoarthritis (KOA).Method Sixty KOA patients were randomized into a treatment group and a control group by using random number table, 30 cases each. The control group was intervened by oral administration of Celecoxib capsules, while the treatment group was given EA, 14 d as a treatment course. The changes of relevant cytokines [apelin, tumor necrosis factor (TNF)-α, TNF soluble receptor (TNFsR)-Ⅰ, TNFsR-Ⅱ, interleukin (IL)-1β, and IL-6] in serum of the two groups were observed.Result The intra-group comparisons of the total score, and the scores of pain, stiffness and dysfunction of the Western Ontario and McMaster Universities (WOMAC) osteoarthritis index showed significant differences in both groups (P<0.05); there were significant between-group differences in comparing the total score, and the scores of pain, stiffness and dysfunction of the WOMAC index after the treatment (P<0.05). The Visual Analogue Scale (VAS) scores were changed significantly after the intervention in both groups (P<0.05); there was nosignificant difference in comparing the VAS score between the two groups after the treatment (P>0.05). The levels of IL-6, TNF-α and TNFsR-Ⅰ were significantly changed after the treatment in both groups (P<0.05); the level of IL-1β was markedly changed after the intervention in the control group (P<0.05); there was a significant change in the level of apelin after the intervention in the treatment group (P<0.05), and there was a significant difference in comparing the level of apelin between the two groups after the treatment (P<0.05).Conclusion EA can produce a satisfactory efficacy in treating KOA; it can significantly improve the symptoms and signs, and mitigate pain and symptoms through regulating the expressions of cytokines.
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Objective To investigate the situation of admission nursing evaluation on elderly patients of geriatrics departments in several Shanghai comprehensive hospitals, and the results will be the base for designing elderly patient′s comprehensive admission nursing evaluation form. Methods 41 nurse managers of geriatrics department in Shanghai comprehensive hospitals were selected as research object. Self-designed questionnaire included general data, elderly patient admission nursing evaluation content and the importance of each item. Results The effective recovery rate was 97.56%(40/41). 98%(39/40) hospitals considered that elderly patients need to have a comprehensive assessment according to the elderly characteristics, only 18%(7/40) hospitals used the special nursing evaluation form for elderly patients. The score of importance of five dimensions were as follows: geriatric syndromes or problems scored (4.64 ± 0.38), physical function assessment scored (4.60 ± 0.46), physiological function assessment scored (4.53 ± 0.45), mental and psychological assessment scored (4.10 ± 0.57) and social assessment scored (4.07±0.52). The score of importance of 29 items were as follows:fall assessment scored (4.90±0.30), pressure ulcer assessment scored (4.85 ± 0.36), activities of daily living scored (4.75 ± 0.44), urinary incontinence assessment scored (4.70 ± 0.52), pain assessment scored (4.70 ± 0.46). Conclusions It is of great significance to carry on the comprehensive admission nursing evaluation for the elderly patients, which needs to be valued and strengthened.
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Background:Intestinal metaplasia(IM)and dysplasia in Barrett’s esophagus(BE)can be detected by endoscopic biopsy combined with pathological examination. Aims:To evaluate the efficacy of acetic acid stain-guided biopsies in detection of IM and dysplasia in BE under standard endoscopy. Methods:One hundred patients found to have columnar-lined esophagus by endoscopy from January 2012 to November 2014 at Shanghai Jing’an District Central Hospital were enrolled. Under standard endoscopy,acetic acid was sprayed and multi-point biopsies were performed in visible abnormal regions. This was followed by four-quadrant biopsies at every 1 to 2 cm of the remaining BE segments. Detection rates of columnar epithelium,IM and dysplasia were compared between acetic acid stain-guided biopsies and four-quadrant biopsies. Results:A total of 96 patients were diagnosed as BE. Detection rates of columnar epithelium by acetic acid stain-guided biopsies and four-quadrant biopsies were 96. 9%(93 / 96)and 90. 6% (87 / 96),respectively,no significant difference was found between the two methods(McNemar test,P = 0. 146),however,consistency of the two methods was poor(κ = 0. 340). A total of 32 patients were diagnosed as BE with IM. Detection rates of IM by acetic acid stain-guided biopsies and four-quadrant biopsies were 84. 4%(27 / 32)and 40. 6%(13 / 32),respectively,difference in detection rate between the two methods was significant(McNemar test,P = 0. 007),and consistency of the two methods was poor(κ =0. 266). Five cases of low-grade dysplasia were detected by acetic acid stain-guided biopsies only. Conclusions:Acetic acid stain-guided biopsies can detect BE with IM and dysplasia better than four-quadrant biopsies.
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<p><b>BACKGROUND</b>Immune cells within a tumor microenvironment have shown modulatory effects on tumor angiogenic activity. Renal cell carcinoma (RCC) is a hypervascular tumor that reportedly increases the frequency of regulatory T cells (Tregs) in tumor tissues. This study investigated the correlation between Tregs infiltration and angiogenic status in RCC.</p><p><b>METHODS</b>Thirty-six patients with RCC were enrolled in the present study, and twenty age-matched healthy donors were included as the control. Tregs were defined as CD4(+)CD25(high)CD127(low/-) T cells. The frequency of Tregs in peripheral blood and tumor infiltrating lymphocytes (TILs) were determined by flow cytometry. The expression of vascular endothelial growth factor (VEGF) in surgical resection specimens were measured with a commercial enzyme-linked immunosorbent assay (ELISA) kit. Microvessel density (MVD) was calculated on slides stained with CD34 antibody. Spearman's rank correlation was performed to evaluate the correlation between the frequencies of Tregs in TILs and VEGF values, as well as between frequencies of Tregs and MVD determinations.</p><p><b>RESULTS</b>Compared to healthy controls, the frequency of peripheral blood Tregs was significantly increased in patients with RCC (P < 0.05). The percentage of tumor-infiltrating Tregs was higher than that of peripheral blood Tregs in patients with RCC (P < 0.01). In addition, the frequency of tumor-infiltrating Tregs was shown to significantly correlate with the pathological stage (P < 0.05) and nuclear grade (P < 0.01). Importantly, a significant positive correlation was observed between the frequency of tumor-infiltrating Tregs and VEGF protein expression (r = 0.51, P < 0.05), as well as between frequencies of Tregs and MVD score (r = 0.39, P < 0.05).</p><p><b>CONCLUSIONS</b>These observations suggest that the high pro-angiogenic status of RCC may be associated with the accumulation of Tregs in the local microenvironment. Angiogenesis networks may be connected with immune tolerance units and cooperate with each other to facilitate tumor growth and progression.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Renal Cell , Allergy and Immunology , Metabolism , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Immunohistochemistry , Kidney Neoplasms , Allergy and Immunology , Metabolism , Lymphocytes, Tumor-Infiltrating , Allergy and Immunology , Neovascularization, Pathologic , Allergy and Immunology , Metabolism , T-Lymphocytes, Regulatory , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To compare the mechanical properties of different dental optimal material selection for orthodontic appliance.</p><p><b>METHODS</b>Four commercialized thermoplastic products under different test conditions, and provide the suggestion of thermoplastic products were tested. The tear strength, elongation at break and stress relaxation of these materials were measured under different test conditions.</p><p><b>RESULTS</b>The tear strength declined after thermoforming, and rose again after 2 weeks of distilled water immersion. The elongation at break rose after thermoforming, and declined after 2 weeks of distilled water immersion. No significant changes were observed for brand A under different test conditions. Brand A showed the slowest stress relaxation of 0.0148 N/s.</p><p><b>CONCLUSIONS</b>The mechanical properties of thermoplastic materials were influenced by environmental factors. Brand A exhibited optimal comprehensive properties.</p>
Subject(s)
Analysis of Variance , Dental Materials , Chemistry , Hot Temperature , Materials Testing , Orthodontic Appliances , Polyethylenes , Chemistry , Polyvinyls , Chemistry , Shear Strength , Stress, MechanicalABSTRACT
ObjectiveTo explore the role of fatty acid on endoplasmic reticulum (ER) stress and to detect the influence of tauroursodeoxycholic acid (TUDCA) on the expression of ER related genes in steatosis hepatocytes by establishing different models of hepatic steatosis.MethodsHealthy adult hepatocyte cell line HL-7702 was taken as research object.The model of hepatic steatosis was established with palmitic acid alone or mixed fatty acids.The cell viability was measured and calculated through CCK-8 positive cell proliferation assay.The cell morphology and steatosis was observed.The intracellular triglyceride content was tested with the triglyceride determination kit.With TUDCA intervention,the relative expression of Glucose-regulated protein 78 (GRP78) and CCAAT/Enhancer binding protein homologous protein (CHOP) at mRNA level was determined by real-time PCR.ResultsThe viability of hepatocyte was influenced once the concentration of mixed fatty acid reached 0.5 mmol/L or palmitic acid was 0.125 mmol/L.The effect of palmitic acid alone was stronger than that of mixed fatty acid on hepatocyte injury.The content of intra-hepatocyte triglyceride gradually increased in mixed fatty acid group,which only significantly increased after treated for 12 hours in palmitic acid alone group and then there was no significant change.There was no significant difference in relative expression of GRP78 and CHOP at mRNA level in various concentrations and treated time of mixed fatty acid group.After treated with palmitic acid at 0.5 mmol/L,intra-hepatocyte relative expression of CHOP at mRNA level increased obviously,however there was no effect on GRP78mRNA expression.After treated with palmitic acid at 1.0 mmol/L,both intra-hepatocyte GRP78 and CHOP mRNA relative expression increased.There was no significant difference in GRP78 and CHOP mRNA relative expression before and after ER stress in TUDCA intervented low dose palmitic acid group.There was significant difference in CHOP mRNA relative expression before and after ER stress in TUDCA intervented high dose palmitic acid group (at 12hrs:8.6400 to 5.1032; at 24hrs:13.7948to 6.4928,P=0.042 and 0.017),while no significant difference in GRP78 mRNA expression.ConclusionAt same fatty acid concentration,the larger propotion palmitic acid has,the more severe injury hepatocyte has.The regulation of palmitic acid on intra-hepatic ER stress is a time and dose dependent manner.TUDCA may improve palmitic acid induced ER stress to some degree.
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<p><b>OBJECTIVE</b>To study the degree and changes of cervical spinal canal stenosis by MRI scans in flexion and extension of the cervical vertebrae.</p><p><b>METHODS</b>Thirty cases of cervical stenosis included 13 male and 17 female with an average age of 39 years ranging from 28 to 66 years. The sagittal diameter of cervical spinal canal were below 10 mm (absolute stenosis) in 12 cases,within 10 to 12 mm (correspondence stenosis) in 18 cases. MRI scans in neutrality, flexion, extension performanced and the degree of cervical spinal canal stenosis and the changes of spinal cord compression were evaluated after MRI scans obtained.</p><p><b>RESULTS</b>Nineteen patients of extension occurrenced stenosis more serious, 8 patients of flexion occurrenced (P < 0.05).</p><p><b>CONCLUSION</b>For the cervical stenosis imaging diagnostic, flexion and extension of cervical MRI scan can be used to supplement conventional MRI examination neutral position, and the extension of MRI is more sensitivity than neutral position and flexion bit.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Cervical Vertebrae , Diagnostic Imaging , Magnetic Resonance Imaging , Methods , Radiography , Spinal Stenosis , Diagnosis , Diagnostic ImagingABSTRACT
<p><b>OBJECTIVE</b>To examine the correlation between the mRNA and proteins expressions of gastrin(GAS), and the association of protein expression of GAS with apoptosis index(AI) and apoptosis regulation gene Fas/FasL, caspases in colorectal cancer.</p><p><b>METHODS</b>The expressions of GAS mRNA in tumor tissues of 79 cases with colorectal cancer were detected by nested RT-PCR. Cell apoptosis was detected by molecular biology in situ apoptosis detecting technic(TUNEL). Protein expressions of GAS, Fas/FasL, and caspases were detected by immunohistochemical staining (SP method).</p><p><b>RESULTS</b>The positive correlation was found between the mRNA and proteins expressions of GAS(rGAS=0.99, P<0.01). The mRNA and protein expressions of GAS in well and moderately differentiated cancers were significantly lower than those in poorly differentiated cancers (chi(2)(high vs low)=10.47, 10.23, P<0.01, chi(2)(middle vs low)=6.68, 4.95, P<0.05). The mRNA and protein expressions of GAS in papillary and tubular adenocarcinomas were significantly lower than those in mucinous adenocarcinomas, signet-ring cell carcinoma and undifferentiated carcinoma (chi(2)(papillary vs mucinous and signet-ring)=4.80, 6.22, chi(2)(papillary vs undifferentiation)=5.44, 8.43, chi(2)(tubular vs mucinous and signet-ring)=4.40, 4.38, chi(2)(tubular vs undifferentiation)=4.92, 6.43, P<0.05, respectively). The mRNA and protein expressions of GAS in Dukes' stages A, B were significantly lower than those in Dukes stages C, D (chi(2)=4.84, 4.45, P<0.01). The AI in GAS high and moderate expression groups of colorectal cancer were significantly lower than that in low expression group (q(high vs low)=6.71, q(middle vs low)=4.60, P<0.01). The positive expression rate of FasL was significantly different among GAS high, moderate and low expression groups of colorectal cancer (chi(2)=9.35, P<0.01). The positive expression rate of FasL in GAS high and moderate expression groups was higher than that in low expression group (chi(2)high vs low=6.24, chi(2)(middle vs low)=4.74, P<0.05).</p><p><b>CONCLUSIONS</b>GAS plays an important role in the regulation of cell apoptosis in colorectal carcinoma, whose mechanism may be related to the aberrant expression of Fas/FasL. GAS will be one of the indicators of the biological behavior in colorectal carcinoma.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Caspases , Metabolism , Colorectal Neoplasms , Metabolism , Pathology , Fas Ligand Protein , Metabolism , Gastrins , Metabolism , RNA, Messenger , Metabolism , fas Receptor , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To scan for mutations of polycystic kidney disease 1 gene (PKD1) in Chinese population in order to find some features about Chinese patients and a better approach to detect mutations.</p><p><b>METHODS</b>Twenty-five PKD-affected individuals from twenty-one unrelated genealogies and sixteen controls participated in the study. Thirty-five blood samples and six tissues were obtained after receiving informed consent and were in accordance with institutional ethical guidelines. Genomic DNA was isolated from peripheral blood using standard procedures. PCR amplification of genomic DNA was performed to generate the aimed fragments. Amplified fragments were analyzed by denaturing gradient gel electrophoresis (DGGE). A GC clamp was attached to the 5' primer. After that, the abnormal fragments were sequenced on freshly amplified specific PCR products with the dideoxynucleotide chain termination method. Sequencing was performed for all samples to evaluate DGGE.</p><p><b>RESULTS</b>Aimed fragments of exons 44 and 45 were amplified. DGGE detected eleven abnormal PCR fragments. Two novel mutations were identified by sequencing, included one nonsense mutation (C12217T) and one frameshift (12431delCT). In addition, one polymorphism (A50747C) was identified. The mutation detection rate is 8% in our study.</p><p><b>CONCLUSION</b>Two novel pathogenic mutations were detected, including one nonsense mutation (C12217T) and one frameshift (12431delCT).</p>