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Objective To establish and forensically verify a 42 microhaplotypes(mircohaps,MHs)mul-tiplex assay system based on next-generation sequencing(NGS),and to explore the application value of this system in the practice of forensic genetics.Methods A total of 42 highly polymorphic MHs were selected from previous studies,and sequenced by the MiSeq FGxTM platform to verify the repeata-bility,sensitivity,specificity,stability,and mixture analysis ability of the detection system.Through population genetic investigation of 102 unrelated Chinese Han individuals in Liyang City,Jiangsu Province,China,the application value of this system in forensic genetics was evaluated.Results The sequencing repeatability of the 42-plex MHs assay was 100%and the sensitivity was as low as 0.062 5 ng.The system had the ability to withstand the interference of indigo(≤2 500 ng/μL),humic acid(≤9 ng/μL),hemoglobin(≤20 μmol),and urea(≤200 ng/μL)and to detect mixtures of 2 people(1∶19),3 people(1∶1∶9)and 4 people(1∶1∶1∶9).Based on 102 individual data,the combined power of discrimination and the combined power of exclusion were 1-3.45×10-30 and 1-3.77×10-11,respectively,and the average effect value of alleles was 2.899.Conclusion The 42-plex MHs assay was successfully established in this study and this system has high repeatability and sensitivity,good anti-jamming ability and mixture analysis ability.The 42 MHs are highly polymorphism and have good application value in individual identification and paternity testing.
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OBJECTIVE@#To explore the effects of prognostic nutritional index (PNI) combined with D-dimer on the prognosis of patients with newly diagnosed diffuse large B-cell lymphoma (DLBCL).@*METHODS@#The clinical data of 73 DLBCL patients at initial diagnosis were retrospectively evaluated, and the optimal cut-off point of PNI and D-dimer were determined by ROC curve. The overall survival (OS) rate and progression-free survival (PFS) rate in different subgroups were compared using Kaplan-Meier survival curves. Univariate and multivariate Cox regression analysis was performed to identify the factors associated with OS.@*RESULTS@#Compared with the low PNI group (PNI<44.775), the high PNI group (PNI≥44.775) had better OS (P =0.022) and PFS (P =0.029), the 2-year OS rates of the two groups were 55.6% and 78.3% respectively (P =0.041). Compared with the high D-dimer group (D-dimer≥0.835), the low D-dimer group (D-dimer<0.835) had better OS (P <0.001) and PFS (P <0.001), the 2-year OS rates of the two groups were 51.4% and 86.8% respectively (P =0.001). Meanwhile, patients in the high PNI+ low D-dimer group had better OS (P =0.003) and PFS (P <0.001) than the other three groups, the 2-year OS rate was statistically different from the other three groups (P <0.05). The multivariate analysis revealed that NCCN-IPI (HR =2.083, 95%CI : 1.034-4.196, P =0.040), PNI (HR =0.267, 95%CI : 0.076-0.940, P =0.040) and PNI+D-dimer (HR =9.082, 95%CI : 1.329-62.079, P =0.024) were the independent risk factors affecting OS in patients with DLBCL. Subgroup analysis showed that PNI, D-dimer, and PNI combined with D-dimer could improve the prognostic stratification in low and low-intermediate risk DLBCL patients.@*CONCLUSION@#High PNI, low D-dimer and combination of high PNI and low D-dimer at initial diagnosis suggest a better prognosis in DLBCL patients.
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Humans , Prognosis , Nutrition Assessment , Retrospective Studies , Lymphoma, Large B-Cell, Diffuse/pathologyABSTRACT
Vascular wall-resident stem cells (VW-SCs) play a critical role in maintaining normal vascular function and regulating vascular repair. Understanding the basic functional characteristics of the VW-SCs will facilitate the study of their regulation and potential therapeutic applications. The aim of this study was to establish a stable method for the isolation, culture, and validation of the CD34+ VW-SCs from mice, and to provide abundant and reliable cell sources for further study of the mechanisms involved in proliferation, migration and differentiation of the VW-SCs under various physiological and pathological conditions. The vascular wall cells of mouse aortic adventitia and mesenteric artery were obtained by the method of tissue block attachment and purified by magnetic microbead sorting and flow cytometry to obtain the CD34+ VW-SCs. Cell immunofluorescence staining was performed to detect the stem cell markers (CD34, Flk-1, c-kit, Sca-1), smooth muscle markers (SM22, SM MHC), endothelial marker (CD31), and intranuclear division proliferation-related protein (Ki-67). To verify the multipotency of the isolated CD34+ VW-SCs, endothelial differentiation medium EBM-2 and fibroblast differentiation medium FM-2 were used. After culture for 7 days and 3 days respectively, endothelial cell markers and fibroblast markers of the differentiated cells were evaluated by immunofluorescence staining and q-PCR. Furthermore, the intracellular Ca2+ release and extracellular Ca2+ entry signaling were evaluated by TILLvisION system in Fura-2/AM loaded cells. The results showed that: (1) High purity (more than 90%) CD34+ VW-SCs from aortic adventitia and mesenteric artery of mice were harvested by means of tissue block attachment method and magnetic microbead sorting; (2) CD34+ VW-SCs were able to differentiate into endothelial cells and fibroblasts in vitro; (3) Caffeine and ATP significantly activated intracellular Ca2+ release from endoplasmic reticulum of CD34+ VW-SCs. Store-operated Ca2+ entry (SOCE) was activated by using thapsigargin (TG) applied in Ca2+-free/Ca2+ reintroduction protocol. This study successfully established a stable and efficient method for isolation, culture and validation of the CD34+ VW-SCs from mice, which provides an ideal VW-SCs sources for the further study of cardiovascular diseases.
Subject(s)
Mice , Animals , Endothelial Cells , Cell Differentiation/physiology , Stem Cells , Adventitia , Fibroblasts , Cells, Cultured , Antigens, CD34/metabolismABSTRACT
ObjectiveTo observe the effects of modified Chaihu Shugansan(CHSG) and its disassembled formulas on angiotensin-converting enzyme 2 (ACE2)-angiotensin (Ⅰ-Ⅶ) [Ang (Ⅰ-Ⅶ)]-mitochondrial assembly receptor (MasR) axis in hyperlipidemic rats with myocardial ischemia and depression, and to explore the underlying mechanism of its prevention and treatment of myocardial ischemia and depression. MethodA total of 108 male SD rats were randomly divided into a normal group, a model group, a modified CHSG group (11.7 g·kg-1), a Quyu Huatan disassembled formula group (4.05 g·kg-1), a Shugan Xingqi disassembled formula group (3.15 g·kg-1), a Jianpi Yangxue disassembled formula group (4.5 g·kg-1), a fluoxetine group (0.001 8 g·kg-1), a trimetazidine group (0.005 4 g·kg-1), and a simvastatin group (0.001 8 g·kg-1), with 12 rats in each group. The hyperlipidemia model with myocardial ischemia and depression was induced with a high-fat diet combined with injection of isoproterenol (ISO) and chronic unpredictable mild stress (CUMS) in rats in the model group and groups with drug intervention for eight weeks. The rats in each group with drug intervention were treated correspondingly by gavage from the first day of modeling, while those in the normal group and the model group received the same amount of normal saline. The behavioral changes of rats in each group were observed by open field test and forced swimming test. Left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) were measured by echocardiography. The serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were detected by the enzyme-labeled apparatus. Hematoxylin-eosin (HE) staining was used to observe the histomorphological changes of the heart. The serum levels of angiotensin Ⅱ (AngⅡ), ACE2, and Ang(Ⅰ-Ⅶ) were detected by enzyme-linked immunosorbent assay (ELISA). The protein and mRNA expression of ACE2 and MasR in the hippocampus and the heart was detected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot. ResultCompared with the normal group, the model group showed reduced movement time, distance, and average speed in the central area of the open field (P<0.01), prolonged immobility time of rats in the forced swimming test (P<0.01), decreased LVFS and LVEF (P<0.01), inflammatory exudation and disorderly arranged fiber in heart tissues, elevated serum levels of TC, LDL-C, AngⅡ, ACE2 and Ang(Ⅰ-Ⅶ), diminished HDL-C (P<0.01), dwindled mRNA and protein expression of ACE2 in the hippocampus and the heart and MasR in the hippocampus, and up-regulated mRNA and protein expression of MasR in the heart (P<0.01). Compared with the model group, the modified CHSG group displayed increased movement time, distance, and average speed in the center area of the open field (P<0.01), shortened immobility time in the forced swimming test (P<0.01), increased LVFS and LVEF (P<0.01), relieved heart injury, reduced serum levels of TC, LDL-C, AngⅡ, ACE2, and Ang(Ⅰ-Ⅶ), elevated level of HDL-C (P<0.01), up-regulated mRNA and protein expression of ACE2 in the hippocampus and the heart and MasR in the hippocampus, and down-regulated mRNA and protein expression of MasR in the heart (P<0.01). Each disassembled formula could improve the above indexes to a certain extent (P<0.05, P<0.01), but the effect of the whole formula was optimal. ConclusionThe modified CHSG and its disassembled formulas have the effects of resisting depression, improving myocardial injury, and reducing blood lipid. Due to the synergistic effects of stasis-resolving/phlegm-eliminating drugs, liver-smoothing/Qi-moving drugs, and spleen-tonifying/blood-nourishing drugs in the formula, the modified CHSG is superior to each disassembled formula in efficacy. Its mechanism may be related to the activation of the ACE2-Ang (Ⅰ-Ⅶ)-MasR axis.
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OBJECTIVE@#To explore the influence of controlling nutritional status (CONUT) score on the prognosis of newly diagnosed patients with multiple myeloma (MM).@*METHODS@#The clinical data 119 patients with MM who were diagnosed according to the international myeloma diagnostic criteria in Lanzhou University Second Hospital from April 2010 to October 2018 were collected and retrospectively analyzed. The relationship between clinical indexes, including age, sex, MM type, absolute lymphocyte count (ALC), absolute neutrophil count (ANC), absolute monocyte count (AMC), hemoglobin (Hb), platelet (PLT), β@*RESULTS@#Compared with high-scoring group, low-scoring group had a better OS [median OS was 43.3 months and 127.67 months, respectively, 95% confidence interval (CI): 57.065-78.345, P=0.038]. At the same time, the low-scoring group also had higher level of ALC, ANC, AMC, Hb, PLT, ALB, and CHO but lower of GLO. Multivariate survival analysis showed that age (HR=1.027, 95%CI: 1.000-1.054, P=0.048), AMC (HR=11.284, 95%CI: 22.968-42.897, P<0.001), CONUT score (HR=1.198, 95%CI: 1.036-1.385, P=0.015), M protein (non-IgG/IgG type) type (HR=0.503, 95%CI: 0.259-0.977, P=0.043) were independent factors affecting the prognosis of MM patients.@*CONCLUSION@#The CONUT score as an immune-nutrition score is a convenient and easy-to-obtain index to effectively predict the prognosis of MM patients.
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Humans , Lymphocyte Count , Multiple Myeloma/diagnosis , Nutritional Status , Prognosis , Retrospective StudiesABSTRACT
Hepatocellular carcinoma (HCC) is the sixth most common malignancy and the fourth leading cause of cancer related death worldwide. China covers over half of cases, leading HCC to be a vital threaten to public health. Despite advances in diagnosis and treatments, high recurrence rate remains a major obstacle in HCC management. Multi-omics currently facilitates surveillance, precise diagnosis, and personalized treatment decision making in clinical setting. Non-invasive radiomics utilizes preoperative radiological imaging to reflect subtle pixel-level pattern changes that correlate to specific clinical outcomes. Radiomics has been widely used in histopathological diagnosis prediction, treatment response evaluation, and prognosis prediction. High-throughput sequencing and gene expression profiling enabled genomics and proteomics to identify distinct transcriptomic subclasses and recurrent genetic alterations in HCC, which would reveal the complex multistep process of the pathophysiology. The accumulation of big medical data and the development of artificial intelligence techniques are providing new insights for our better understanding of the mechanism of HCC via multi-omics, and show potential to convert surgical/intervention treatment into an antitumorigenic one, which would greatly advance precision medicine in HCC management.
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Humans , Artificial Intelligence , Carcinoma, Hepatocellular/therapy , Gene Expression Profiling , Liver Neoplasms/genetics , PrognosisABSTRACT
OBJECTIVE@#To investigate the value of red blood cell distribution width (RDW) and fibrinogen (Fib) level for the evaluation of therapeutic efficacy and prognosis in patients with diffuse large B-cell lymphoma (DLBCL).@*METHODS@#The relationship between RDW/Fib at initial diagnosis and efficacy and the clinical outcome was retro-spectively analyzed based on the study of 105 patients with DLBCL. The patients were divided into two groups: low RDW group (≤15%) and high RDW group (>15%), low Fib group (Fib≤4 g/L) and high Fib group (Fib>4 g/L) according to the normal values of RDW and Fib. Therapeutic efficacy, overall survival (OS) time and progression free survival (PFS) time were compared between two groups. The correlation between each factors and efficacy, prognosis was analyzed by univariate and multivariate regression.@*RESULTS@#The therapeutic efficacy (P<0.001), OS time(P=0.004), and PFS time(P=0.007) were poorer in the high RDW group as compared with the low RDW group. The efficacy (P=0.015) and PFS time(P=0.04) were poorer in the high Fib group as compared with the low Fib group. Multivariate analysis showed that high RDW was the independent risk factor for efficacy of DLBCL patients (OR=3.394, 95% CI 1.093-10.539, P=0.035).@*CONCLUSION@#High RDW and high Fib associate with poor efficacy in DLBCL patients.
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Objective The imbalance of glucagon secretion plays an important role in the development of diabetes. The newly discovered ACE2/Ang(1-7)/Mas pathway is an vital branch of the RAS system and has regulatory effects on islet function, but its effect on glucagon secretion is still unknown. The article aimed to investigate the effect and possible mechanisms of AVE0991, a Mas receptor agonist on glucagon secretion of diabetic db/db mice. Methods A tolal of 30 db/db mice were randomized to AVE group and model group (n=15), and their age-matched nondiabetic db/m mice were selected as the normal group (n=15). The mice in AVE group were treated with AVE0991 20mg/kg/d and those in model group were treated with placebo via gavage for 6 weeks. The metabolic indicators were measured every week. After 6 weeks of treatment, intraperitoneal glucose tolerance test (IPGTT) and islet perifusion were performed to evaluate glucagon release kinetics in vivo and vitro. Double-label immunofluoresence assay was adoppted to assess the content of α and β cells. Moreover, qRT-PCR and western blot were employed to detect the GCK expression in islets. Results The fasting blood sugar[(19.1±0.8)mmol/L] and glucose tolerance [(14.1±0.5) mU/L]of AVE group were significantly lower than those of the model group[(25.3±3.0)mmol/L,(17.3±1.8)mU/L](P<0.05) and still higher than those of normal group[(6.3±0.5)mmol/L,(5.7±0.3)mU/L](P<0.05). At the 30, 60, and 120 min after IPGTT, the blood glucose level and glucagon level in AVE group were lower than model group, but still higher than the normal group (P<0.05). During low glucose perfusion, the glucagon secretion level of the islets of normal group [(20.6±0.5 pmol/L)] was lower than that of model group [(29.1±0.7) pmol/L)] and AVE group [(27.6±0.8) Pmol/L], and the difference was statistically significant (P<0.05). During high glucose perfusion, there was a statistically significant difference in glucagon level between AVE group and model group at 30 and 60 min(P<0.05). Semi-quantitative analysis showed that the islet α-cell content of model group [(3.3±0.7) mg] was significantly higher than that of normal group [(1.2±0.3) mg] (P<0.05), and the β cell content [(2.4±0.6) mg] was significantly lower than that of normal group [(4.8±0.3) mg] (P<0.05); While compared with the model group, the islet α-cell content [(1.8±0.4) mg] decreased significantly (P<0.05) and the β-cell content [(4.2±0.5) mg] increased significantly in AVE group (P<0.05). The expression levels of GCK mRNA and protein in model group were lower than those in normal group (P<0.05). Conclusion Activation of Mas receptors can improve glucose metabolism by reducing the secretion of glucagon after glucose load in db/db diabetic mice. The mechanism may be related to the decrease of islet α cell content and the increase of islet GCK expression.
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Objective: To investigate the effect of Erzhiwan (EZW) on the invasion and metastasis of human colon cancer HCT116 cells. Method: The abilities of invasion (number of transmembrane cells) and migration (relative width of 48 h scratch) were observed by Transwell assay. Western blot was used to detect vascular endothelial growth factor (VEGF) and E-cadherin protein, respectively. Result: ① Transwell results showed that compared with the blank control group, the number of model piercing cells in each drug group was decreased (PPPPPPPP-1 L-OHP combined with 10% EZW group was lower than that in L-OHP group (PP-1 L-OHP combined with 10% EZW group and L-OHP group increased the expression of E-cadherin protein significantly. Compared with L-OHP group, the expression of E-cadherin protein was the highest (PConclusion: EZW can inhibit the invasion and metastasis of colon cancer HCT116 cells, and the best effect is achieved after combined with L-OHP. This may be related to the decrease of VEGF protein expression and the increase of E-cadherin protein expression after combination with L-OHP.
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Aim To study the effect of tetrandrine ( Tet ) on proliferation of MCF-7 breast cancer cells and the possible mechanism underlying this biological process. Methods CCK-8, flow cytometric and Western blot were introduced to analyze the effect of Tet on proliferation and apoptosis in MCF-7 cells.Re-al-time PCR and/or Western blot assay were employed to detect the effect of Tet on expression of IGFBP-5 , p53 and MDM2.CCK-8 and recombinant adenovirus were utilized to determine the effect of IGFBP-5 on the proliferation inhibitory effect of Tet .Western blot assay was introduced to evaluate the effect of IGFBP-5 on p53 which was induced by Tet .Results Tet inhibited the proliferation , arrested cell cycle at G 1 phase and decreased the expression of PCNA concentration dependently in MCF-7 cells.Meanwhile, Tet increased the percentage of apoptotic cells , the level of Bad and reduced the level of Bcl-2.Tet increased the expres-sion of IGFBP-5 either mRNA or protein , over-expres-sion of IGFBP-5 enhanced the anti-proliferation activity of Tet in MCF-7 cells, but knockdown of IGFBP-5 at-tenuated this effect of Tet .Tet increased the level of p53 and decreased that of MDM2, and exogenous IG-FBP-5 enhanced the effect of Tet on p53 and MDM2, respectively .Conclusion Tet can inhibit the prolifer-ation of MCF-7 cells, and this activity is partly media-ted by increasing the function of p 53 signal , which may be triggered by the Tet-induced IGFBP-5.
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Objective The aim of this study was to assess the effect of hemodialysis combined with hemoperfusion for the treatment of microinflammation in diabetic nephropathy patients.Methods One hundred and twenty-five cases of diabetic nephropathy experienced treatment from July 2014 to December 2015 in author's hospital were analyzed retrospectively.They were divided into two groups randomly.The observation group received hemodialysis combined with hemoperfusion,and the control group received just hemodialysis therapy.The status of microinflammation and curative effect were compared.Results The effective rate of observation group was 60.6%,while was 45.8% in control group (P<0.05).Microinflammation level:compared with control group,SAA,IL-6,IL-17,IL-22,IL-1β,TNF-αand TGF-β1 status of observation group showed decreased trend (P<0.01).Conclusion Hemodialysis combined with hemoperfusion can effectively decreased microinflammation status in diabetic nephropathy.
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This study was aimed to establish a method to create a stable planar lipid bilayer membranes (PLBMs), in which large conductance calcium-activated potassium channels (BK) were reconstituted. Using spreading method, PLBMs were prepared by decane lipid fluid consisting of N-weathered mixture of phosphatidylcholine and cholesterol at 3:1 ratio. After successful incorporation of BKchannel into PLBMs, single channel characteristics of BKwere studied by patch clamp method. The results showed that i) the single channel conductance of BKwas (206.8 ± 16.9) pS; ii) the activities of BKchannel were voltage dependent; iii) in the bath solution without Ca, there was almost no BKchannel activities regardless of under hyperpolarization or repolarization conditions; iv) under the condition of +40 mV membrane potential, BKchannels were activated in a Caconcentration dependent manner; v) when [Ca] was increased from 1 μmol/L to 100 μmol/L, both the channel open probability and the average open time were increased, and the average close time was decreased from (32.2 ± 2.8) ms to (2.1 ± 1.8) ms; vi) the reverse potential of the reconstituted BKwas -30 mV when [K] was at 40/140 mmol/L (Cis/Trans). These results suggest that the spreading method could serve as a new method for preparing PLBMs and the reconstituted BKinto PLBMs showed similar electrophysiological characteristics to natural BKchannels, so the PLBMs with incorporated BKcan be used in the studies of pharmacology and dynamics of BKchannel.
Subject(s)
Animals , Calcium , Chemistry , Electrophysiological Phenomena , Large-Conductance Calcium-Activated Potassium Channels , Chemistry , Lipid Bilayers , Chemistry , Membrane Potentials , Patch-Clamp TechniquesABSTRACT
Objective: To optimize the preparation process of morellic acid B (MAB) mPEG liposomes (MAB-mPEG-LPS), and to study the in vitro release behavior and pharmacokinetics of MAB-mPEG-LPS in rats. Methods: The analytical method of MAB was established; Encapsulation efficiency and particle size were used as the indexes to optimize the mPEG liposomes by orthogonal test, and the highest encapsulation efficiency of MAB-mPEG-LPS was obtained; Transmission electron microscope was used to observe the surface morphology of MAB-mPEG-LPS, and the stability of MAB-mPEG-LPS was measured in 60 d. Dialysis method was also adopted to study the MAB-mPEG-LPS release in vitro; Male SD rats were injected with MAB (1.50 mg/kg), MAB-LPS (1.50 mg/kg), MAB-mPEG-LPS (1.50 mg/kg) via tail vein, and differences in pharmacokinetics parameters of MAB, MAB-LPS, and MAB- mPEG-LPS were compared. Results: The optimized formula of MAB-mPEG-LPS: HSPC was 128 mg, mPEG was 10 mg, HSPC-CHOL was 8∶1. Encapsulation efficiency of MAB-mPEG-LPS was 83.21%. MAB-mPEG-LPS had uniform particle size and smooth surface; In vitro release results showed that the MAB-mPEG-LPS had slow release and long-term effect. It was stable in 60 d; In vivo study showed that t1/2β of MAB in MAB-mPEG-LPS was 66.925 min, which was 4.43 fold to MAB. MAB-mPEG-LPS was 3.29 fold to MA-LPS; AUC0-∞ of MAB in MAB-mPEG-LPS was 241.372 mg∙min/L, which was 3.64 fold to MAB. MAB-mPEG-LPS was 1.99 fold to MAB-LPS. Conclusion: MAB-mPEG-LPS could prolong the circulation time and increase AUC0-∞ of MAB in rats.
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Objective:To compare the safety and effectiveness of two minimally invasive approaches for multi-vessel coronary revascularization.Methods:From August 2014 to February 2017,70 consecutive patients who underwent minimally invasive coronary artery bypass grafting in Peking University Third Hospital were randomly divided into two groups.In one group,40 patients underwent staged-hybrid coronary revascularization (staged-HCR) treatment;in the other group,30 patients underwent minimally invasive total arterial revascularization with bilateral internal thoracic artery (BITA).In staged-HCR group,the patients underwent minimally invasive direct coronary artery bypass grafting (MIDCAB) and percutaneous coronary intervention (PCI) procedure for treatment of multi-vessel disease.In BITA group,the patients underwent total arterial coronary artery bypass grafting with composite "Y" BITA graft.Preoperative and postoperative data of the two groups,including postoperative blood usage,mechanical ventilation time,domiciling duration in intensive care unit (ICU),major adverse cerebral and cardiovascular event (MACCE),and postoperative coronary angiography results were compared,in order to evaluate the safety and effectiveness of these surgical approaches.Results:The preoperative characteristics of 70 patients in the two groups showed no significant difference.All the patients underwent successfully,elective minimally invasive multi-vessel coronary artery bypass grafting as scheduled preoperatively.Postoperative result showed the patients in staged-HCR group took advantages in less postoperative mechanical ventilation time [Staged-HCR group (11.2 ± 8.7) h vs.BITA group (18.3 ± 9.1) h,P =0.013],shorter domiciling duration in ICU [Staged-HCR group (26.29 ± 4.05) h vs.BITA group (44.74 ± 28.75) h,P =0.022],and less total drainage [Staged-HCR group (695.57 ± 250.46) mL vs.BITA group (1 103.26±547.44) mL,P =0.03] than the patients in the group of minimally invasive total arterial revascularization with BITA.Postoperative in hospital coronary angiography showed satisfactory graft patency rates in both groups [97.5% in Staged-HCR group vs.97.8% in BITA group].No MACCE occurred in both groups during hospitalization.Conclusion:Staged-HCR is a feasible method for the treatment of multi-vessel revascularization involving right coronary artery.Minimally coronary revascularization with BITA is associated with superior long-term graft patency and it's recommended for patients who could not tolerate dual-antiplatelet therapy.This study shows that both minimally invasive surgical approaches are safe and effective for treatment of patients with multi-vessel coronary artery disease.
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BACKGROUND: It is still a problem to achieve early and accurate diagnosis of Alzheimer's disease, and delayed treatments often occur in a large number of patients because of late diagnosis or misdiagnosis. Therefore, the development and improvement of related bioanalytical methods are of great importance for the biomarker detection of Alzheimer's disease which is still lack of means that are sensitive, efficient and low-cost. OBJECTIVE: To summarize the diagnostic methods and related biomarkers of Alzheimer's disease, and to sum up the research progress in novel biomimetic receptors for the early detection of Alzheimer's disease biomarkers. METHODS: PubMed, CNKI and Wanfang databases were used to search articles related to the biomarker studies of Alzheimer's disease and relevant studies about methods of biomarkers detection published from 2000 to 2016. The key words were "Alzheimer's disease, biomarkers, detect/detecting/detection" in Chinese and English, respectively. Finally 40 articles were obtained for the review. RESULTS AND CONCLUSION: At present, the bioanalytical methods used for biomarker detection of Alzheimer's disease mostly utilize antibodies as recognition and capture elements of biomarkers, but there are some limitations using traditional antibodies as detection receptors. Thus, novel biomimetic receptors can be substituted for conventional antibodies. Novel biomimetic receptors have high specificity, small size, low production costs and high product stability, and their chemical modification process is relatively convenient. Biomimetic receptors developed for protein analysis include aptamers, polypeptide receptors, peptoid receptors, molecularly imprinted polymers, nanobodies, gelsolin and cucurbit urils. Detection of biomarkers with novel biomimetic receptors instead of conventional antibodies will be more accurate and timely in the early diagnosis of Alzheimer's disease.
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Objective To evaluate the application of novel fluorescent staining method with hypersensitivity and enhancement for microscopic examination of superficial fungal.Methods A total of 200 cases with clinically suspected superficial fungal infection were screened by the hypersensitive,enhanced fluorescent staining,calcofluor white staining and microscopic examination with 1.78 mol/L potassium hydroxide (KOH) based smear assay,respectively.The positive percentages of the different methods were calculated and compared.Results In the developed fluorescent staining examination,clean background was shown in the view field.The hyphae and spores displayed in bright blue,thus fungal morphology was so distinct that they were easy to be distinguished.Compared with calcofluor white staining and KOH based smear,the fluorescent staining showed high positive rate with significant difference respectively (x2 =17.984,P < 0.05;x2 =32.063,P < 0.05).Conclusion The novel fluorescent staining should be a rapid,accurate method for microscopic examination of fungi,which is worthy to be widely used in clinical laboratories for early diagnosis of fungal infection.
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<p><b>OBJECTIVE</b>The device of a bone age film of analysis and process was designed, can calculate the expected patient's height and identify growth level and development.</p><p><b>METHODS</b>Select the DR bone age of 100 children of 6-13 years old, used semi Lagrange algorithm of target region of interest on bone age piece (ROI) for image analysis, compared with 2 pediatric endocrinologists (A, B) who used TW3 artificial to judge bone age (two methods were detected 2 times), and report the results.</p><p><b>RESULTS</b>Bone age assessment process, forecast error of bone age reduced to 0.12 years.</p><p><b>CONCLUSIONS</b>This device can quickly calculate the expected patient's height and identify his growth level, improve the speed and accuracy of bone age assessment, especially in the poor medical conditions in rural and remote areas.</p>
Subject(s)
Adolescent , Child , Humans , Age Determination by Skeleton , Equipment DesignABSTRACT
Spontaneous remission (SR) of leukemia is a rare event in clinic, which possibly correlated with severe infection and sepsis, but its exact mechanism has not been confirmed. Plasmacytoid dendritic cells (pDC) and myeloid dendritic cells (mDC) play a key role in innate and adaptive immunity respectively. A patient with severe infection of staphylococcus aureus acquired completely spontaneous remission (SR), moreover a increased number of pDC were observed, suggesting that bacteria-activated pDC may play an important role in SR. This study was purposed to explore if the bacteria can stimulate pDC successfully and get a functional pDC. Both pDC and mDC were isolated from freshly collected, leukocyte-rich buffy coats from healthy blood donor and leukemic patient with SR by using MACS and FACS. The pDC were cultured in RPMI 1640 medium and were stimulated with different kinds of bacteria and the expression of CD40, CD86 and HLA-DR on the cell surface was analyzed by flow cytometry. The cytokine (IFN-α, IL-12, IFN-γ, IL-2, IL-4, IL-10) production was measured by using ELISA kits. The results showed that the stimulation with staphylococcus aureus and pseudomonas aeruginosa resulted in the maturation of pDC, which secrete a large number of IFN-α and promote the differentiation of naive CD4⁺ T cells to Th1 cells. The activated pDC expressed high level of CD40 and CD86 and showed higher T cell stimulatory capacities. It is concluded that staphylococcus aureus and pseudomonas aeruginosa can activate pDC, the activated pDC secrete high quantity of IFN-α. This result suggests that bacteria stimulated pDC may play a key role in SR of leukemia following severe infections.
Subject(s)
Humans , CD4-Positive T-Lymphocytes , Dendritic Cells , Allergy and Immunology , Microbiology , Interferon-alpha , Interleukin-10 , Interleukin-12 , Interleukin-2 , Interleukin-4 , Leukemia , Diagnosis , Allergy and Immunology , Microbiology , Remission, Spontaneous , Staphylococcus aureusABSTRACT
The aim of present study was to explore the vasodilatation mechanism of angiotensin II (AngII) at the molecular level by investigating the effect of AngII on large-conductance Ca²⁺-activated potassium channels (BK(Ca)) in human mesenteric artery smooth muscle cells. The effect of AngII on BK(Ca) was observed by using patch clamp single channel recording technique and amphotericin-perforated whole-cell recording technique. AngII type 1 receptor (AT₁R) and AngII type 2 receptor (AT₂R) mRNA expression in human mesenteric artery was detected by RT-PCR. In cell-attached patch (Vm = +40 mV), AngII (100 nmol/L) had no significant effect on BK(Ca). After pretreatment with Valsartan (a specific inhibitor of AT₁R, 10 μmol/L), 25, 100 and 250 nmol/L AngII stimulated BK(Ca) activity significantly in a dose response manner. After pretreatment of Valsartan, AngII (100 nmol/L) enhanced BK(Ca) open probability (NP(O)) from 0.010 ± 0.003 to 0.039 ± 0.015, decreased the mean close time (T(C)) of BK(Ca) markedly from (2 729.5 ± 808.6) ms to (487.7 ± 182.5) ms (n = 11, P < 0.05) , but AngII had no significant influences on the amplitude (Amp) and the mean open time (T(O)) of BK(Ca). Further PD123,319 (a specific inhibitor of AT₂R) treatment prevented the stimulatory effect of AngII: PD123,319 decreased the NP(O) of BK(Ca) from 0.016 ± 0.003 to 0.004 ± 0.001 (n = 5, P < 0.05), but had no significant influences on Amp, T(O) and T(C) of BK(Ca). In addition, after pretreatment with Valsartan and PD123,319, AngII (100 nmol/L) had no significant effect on BK(Ca). In the amphotericin-perforated whole-cell patch-clamp configuration, after pretreatment with Valsartan, the current density of BK(Ca) at the voltage of -60 - +30 mV had no significant changes before and after adding 100 nmol/L AngII, but the current density of BK(Ca) at the voltage of +40 mV, +50 mV and +60 mV increased significantly after adding 100 nmol/L AngII, from (9.03 ± 2.23) pA/pF, (12.88 ± 2.55) pA/pF and (17.26 ± 2.84) pA/pF to (12.47 ± 2.22) pA/pF, (18.71 ± 2.51) pA/pF and (27.21 ± 3.12) pA/pF (n = 6, P < 0.05), respectively. Using RT-PCR, the AT₁R mRNA and AT₂R mRNA from isolated human mesenteric artery were detected. So we can draw a conclusion, AngII can stimulate BK(Ca) activity in human mesenteric artery smooth muscle cells after pretreatment with Valsartan, which is possibly mediated by AT₂R.
Subject(s)
Humans , Angiotensin II , Pharmacology , Large-Conductance Calcium-Activated Potassium Channels , Metabolism , Mesenteric Arteries , Cell Biology , Muscle, Smooth, Vascular , Cell Biology , Myocytes, Smooth Muscle , Metabolism , Patch-Clamp Techniques , Receptor, Angiotensin, Type 1 , Metabolism , Receptor, Angiotensin, Type 2 , Metabolism , Tetrazoles , Pharmacology , Valine , Pharmacology , Valsartan , VasodilationABSTRACT
This study was aimed to investigate the immunological effect of modified dendritic cells (DC) which inducing cytotoxic T cells (CTL) against lymphoma cells. The DC were isolated from the lymph node and peripheral blood of patients with diffuse large B cell lymphoma (DLBCL). DC were transfected with recombinant adenovirus vector carrying human p53 gene (rAd-p53-DC). The expression of p53 gene was detected by flow cytometry. Western-blot was used to detect the expression of P53. ELISA was used to detect IL-12 level in supernatant. The mixed lymphocyte reaction (MLR) was used to detect the proliferative ability of auto-lymphocyte stimulated by DC. The lactate dehydrogenase (LDH) release test was used to determine the cytotoxicity of CTL. The results indicates that the expressions of DC surface molecule (except for CD1a) such as CD83, CD80, CD86 and HLA-DR were significantly higher in experiment group than that in control group and blank control group. The secretion of IL-12 in supernatant was higher in experiment group than that in control group. The autologous T lymphocyte proliferation and cytotoxic activity against the same kind of DLBL-cells increased in experiment group as compared with control group and blank control group (P < 0.05). The ability to stimulate T lymphocyte proliferation increased with the rising of the ratio of DC and T lymphocyte. However, there was statistically significant difference between rAd-p53-DC derived from Lymph node and peripheral blood (P < 0.05). It is concluded that rAd-p53-transfected DC can induce CTL response in vitro against lymphoma cells.