Sonoclot analyzer has been widely used in many countries. But the reference intervals provided by the manufacturer were derived from only 45 participants, and there was no cut-off value for transfusion for Sonoclot an...Sonoclot analyzer has been widely used in many countries. But the reference intervals provided by the manufacturer were derived from only 45 participants, and there was no cut-off value for transfusion for Sonoclot analysis. This study aimed to establish reference intervals and transfusion criterion for Sonoclot analysis. Volunteers were recruited from healthy Chinese adults and patients undergoing cardiac surgery. Blood samples were withdrawn from forearm vein and measured for activated clotting time（ACT）, clot rate（CR）, platelet function（PF）, activated partial thromboplastin time（APTT）, fibrinogen concentration（FIB）, and platelet count（PLT）. The reference intervals were determined by the nonparametric method. Cut-off values were determined by the receiver operating characteristics curve. A total of 135 healthy volunteers and 281 patients were enrolled. The 95% reference intervals were 96–195 s, 22–51 signal U/min, 〉1.6 for ACT, CR, PF respectively. In the 281 patients, the results of APTT, FIB, PLT, ACT, CR, and PF ranged from 20.5–300.0 s, 0.28–4.11 g/L,（19.0–387.3）×109/L, 80–514 s, 2.9–74 signal U/min, and 0.1–5.1 respectively. The cut-off values for transfusion were 〉208, ≤14, and ≤1.3 for ACT, CR, PF respectively. The cut-off values of Sonoclot analysis were within the manufacturer＇s reference intervals, while they were outside the reference intervals established in this study. The results suggested that the manufacturer＇s reference intervals were not suitable for Chinese. The reference intervals and cut-off values established in this study will be helpful to Chinese patients.展开更多
Purposes: Hypofibrinogenemia is usually complicated with severe and massive upper gastrointestinal (GI) bleeding, especially in liver cirrhosis. In Japan, neither fibrinogen concentrate nor cryoprecipitate (CRYO) is a...Purposes: Hypofibrinogenemia is usually complicated with severe and massive upper gastrointestinal (GI) bleeding, especially in liver cirrhosis. In Japan, neither fibrinogen concentrate nor cryoprecipitate (CRYO) is available for cases of acquired hypofibrinogenemia to recover the hemostatic level of fibrinogen rapidly. We prepared and produced in-house CRYO from fresh frozen plasma, and compared mortality between pre-implementation and post-implementation of CRYO. Methods: We performed a retrospective cohort study of patients admitted to our single tertiary medical center with upper GI bleeding from January 2011 to December 2016. The observational control period was from January 2011 to February 2013. From March 2013 to December 2016, we implemented the transfusion of CRYO, which was prepared and produced in our hospital. Clinical outcomes were compared between the patients in the two periods. Patients: Eleven patients in the control period and 10 in the intervention period were eligible for analysis. Results: In-hospital mortality (55% vs. 20%, P = 0.238) and mortality within 24 hour after admission (27% vs. 0%, P = 0.246) tended to be lower in the intervention period than in the control period, although the patients had more severe coagulation on admission than those in the control period. The plasma fibrinogen level before the treatment of hemostasis in the intervention period was higher than that in the control period (80 ± 9 mg/dL vs. 127 ± 15 mg/dL, P Conclusion: Implementation of in-house CRYO transfusion may reduce the rate of mortality due to severe upper GI bleeding.展开更多
<strong>Objective:</strong> In Japan, red blood cell (RBC) solution is usually transported by car from a medical institution to the patient’s house for home transfusion. However, there are no regulations ...<strong>Objective:</strong> In Japan, red blood cell (RBC) solution is usually transported by car from a medical institution to the patient’s house for home transfusion. However, there are no regulations for transporting blood by car in the medical setting. Therefore, we assessed and compared the methods (containers) used for transporting the RBC solution by car. <strong>Materials and Methods:</strong> Irradiated RBC solution samples (280 mL) supplied by the Japan Red Cross Society were each divided into two bags. The quality of blood transported by car (1 - 2 hours) in an active transport refrigerator (ATR) (control group) was compared with that transported in a cooler, or styrofoam box (study group). We tested the hemolytic effects of transportation by car, storage, and filtration through a transfusion set on the lactate dehydrogenase (LD) levels in the RBC solutions. <strong>Results:</strong> Post-filtered LD levels were significantly higher in the RBC solutions transported in a cooler-box with inadequate temperature control when compared to those transported in an ATR with optimal temperature control. However, under conditions of optimal temperature control, the post-filtered LD levels were comparable in the control and study (both cooler and styrofoam boxes) group RBC solutions. <strong>Conclusion:</strong> Temperature management is critical for the maintenance of the quality of the RBC solution transported by car.展开更多
Sickle cell disease is an autosomal recessive genetic disease. Vaso occlusive crisis (VOC) is frequently seen in such patients. Painful VOC is usually recurrent, of variable severity due to many factors and its manage...Sickle cell disease is an autosomal recessive genetic disease. Vaso occlusive crisis (VOC) is frequently seen in such patients. Painful VOC is usually recurrent, of variable severity due to many factors and its management poses important challenge in the clinical practice. Few patients do not respond to standard therapies and continue to suffer severe pain for prolonged period or land to serious life threatening situation. The red cell exchange by aphaeresis is presumed to be one efficient alternative in this situation which can reduce the level of HbS below 40% - 50%. However, it is costly and not available everywhere. Both circumstances are common in our state where incidence of sickle cell disease is quite high. In such situations simple red cell exchange i.e. removing 1 unit (350 ml) of blood manually (by phlebotomy) and replacement with one unit normal red cell is effective. All of our four cases of SCA with severe acute VOC, are not responding to standard therapy but responded efficiently to this simple red cell exchange transfusion. Our present observation may pave the way of one simple, affordable, and effective measure to reduce the pain of severe acute VOC not responding to standard therapy. Moderate reduction of HbS by 8% - 14% by simple red cell exchange transfusion was associated with relief of pain of acute VOC;a new observation was reported in all our 4 cases which need to be validated by larger controlled studies.展开更多
BACKGROUND Critically ill patients often present on admission or develop acute respiratory failure requiring intubation and application of positive pressure ventilation during their hospital stay.AIM To investigate an...BACKGROUND Critically ill patients often present on admission or develop acute respiratory failure requiring intubation and application of positive pressure ventilation during their hospital stay.AIM To investigate and identify the epidemiological data,parameters associated with respiratory settings or the mechanics,and values related to arterial blood gases(ABGs)that are associated with outcomes in critically ill patients.METHODS A retrospective analysis of 131 patients[mean age,67.3 years;mean acute physiology and chronic health evaluation(APACHE)score,21.4]with acute respiratory failure requiring invasive mechanical ventilation was performed.The parameters that were statistically analyzed included demographic data,the presence of comorbidities,the presence of coronavirus disease 19(COVID-19),the respiratory rate(RR),peak airway pressure(Ppeak),minute ventilation(MV),positive endexpiratory pressure,and the values related to ABGs.In order to facilitate the statistical analysis,patients were evaluated and compared in groups:Survivors(n=41)vs non-survivors(n=90)and patients without acute kidney injury(AKI)(n=60)vs patients with AKI(n=71).Four endpoints were studied:Mortality,length of stay,duration of mechanical ventilation,and AKI.Group comparisons were performed using the following statistical tests:Theχ^(2) test with Yates’correction,Fisher’s exact test,the Mann-Whitney U test,and Spearman’s rank correlation analysis.Binary logistic regression analysis conducted after the univariate statistical tests facilitated the investigation of the independent predictors of mortality and AKI.A two-sided P value of less than 0.05 was considered the threshold of statistical significance.RESULTS Non-survivors presented statistically significant differences in terms of being older in age,the presence of comorbidities,elevated APACHE score,medical(vs surgical)reasons for admission,presence of COVID-19,lower pH at ABGs,lower values of the oxygenation ratio(arterial oxygen partial pressure to the fraction of inspired oxygen)and arterial oxygen partial pressure,and elevated values of Ppeak,positive end-expiratory pressure,RR,arterial carbon dioxide partial pressure,and MV.The factors identified as independent predictors of mortality were the presence of comorbidities,APACHE score,COVID-19 status,arterial carbon dioxide partial pressure,Ppeak,RR,and MV.COVID-19 presence and elevated values of RR and Ppeak were positively correlated with the other three endpoints(length of stay,the duration of mechanical ventilation in survivors,and the occurrence of AKI in the entire study population)that were studied.The other parameters exhibited a variable(either positive/negative,or no)correlation to the four endpoints under investigation.CONCLUSION Among all investigated outcome measures,COVID-19,Ppeak,and RR were strongly associated with all the endpoints studied,suggesting that proper interventions involving the modifiable respiratory parameters Ppeak and RR could improve the overall outcome in these patients.A novel finding of this study was the relationship between RR and AKI,which is worthy of further investigation.Future studies may explore the clinical interpretation of these findings to improve outcomes in critically ill patients with acute respiratory failure.展开更多
BACKGROUND Low back pain is a major cause of disability worldwide,with intervertebral disc degeneration contributing to nearly 40%of cases.Conventional treatments focus on symptom relief without addressing the underly...BACKGROUND Low back pain is a major cause of disability worldwide,with intervertebral disc degeneration contributing to nearly 40%of cases.Conventional treatments focus on symptom relief without addressing the underlying degeneration.Platelet-rich plasma(PRP),a regenerative therapy rich in growth factors,offers potential therapeutic benefits through growth factor-mediated mechanisms,though clinical evidence is limited.AIM To evaluate the efficacy of intradiscal autologous PRP injection in reducing pain and improving function in patients with chronic lumbar disc prolapse.METHODS This pilot quasi-experimental study was conducted in tertiary care centre between July 2022 and June 2024.The study involved comparing the outcomes between group A(n=17)who failed to respond to conservative treatment measures and received intradiscal PRP injection with group B(n=22)who responded to conservative treatment.Visual Analogue Scale(VAS)and Functional Rating Index(FRI)scores were recorded at baseline,3 weeks,and 6 weeks for both the groups.RESULTS Forty patients were enrolled in the study.The PRP group demonstrated significant improvement in VAS and FRI scores compared to baseline.While both groups improved from their respective baselines,direct between-group comparisons are limited by baseline differences in symptom severity.Patients who failed conservative trial showed significant improvement following PRP intervention,with outcomes approaching those observed in physiotherapy responders.CONCLUSION Intradiscal PRP injection significantly improved pain and function in patients with lumbar disc disease,with clinical improvements that approached the level observed in physiotherapy responders,despite baseline differences in symptom severity.PRP shows promise as an effective treatment for lumbar disc pathology;however,these preliminary findings are limited by the small sample size and short follow-up,warranting larger trials with long-term evaluation.展开更多
Constructing an in vitro vascularized liver tissue model that closely simulates the human liver is crucial for promoting cell proliferation,mimicking physiological heterogeneous structures,and recreating the cellular ...Constructing an in vitro vascularized liver tissue model that closely simulates the human liver is crucial for promoting cell proliferation,mimicking physiological heterogeneous structures,and recreating the cellular microenvironment.However,the layer-by-layer printing method is significantly constrained by the rheological properties of the bioink,making it challenging to form complex three-dimensional vascular structures in low-viscosity soft materials.To overcome this limitation,we developed a cross-linkable biphasic embedding medium by mixing low-viscosity biomaterials with gelatin microgel.This medium possesses yield stress and self-healing properties,facilitating efficient and continuous three-dimensional shaping of sacrificial ink within it.By adjusting the printing speed,we controlled the filament diameter,achieving a range from 250μm to 1000μm,and ensuring precise control over ink deposition locations and filament shapes.Using the in situ endothelialization method,we constructed complex vascular structures and ensured close adhesion between hepatocytes and endothelial cells.In vitro experiments demonstrated that the vascularized liver tissue model exhibited enhanced protein synthesis and metabolic function compared to mixed liver tissue.We also investigated the impact of varying vascular densities on liver tissue function.Transcriptome sequencing revealed that liver tissues with higher vascular density exhibited upregulated gene expression in metabolic and angiogenesis-related pathways.In summary,this method is adaptable to various materials,allowing the rheological properties of the supporting bath and the tissue's porosity to be modified using microgels,thus enabling precise regulation of the liver tissue microenvironment.Additionally,it facilitates the rapid construction of three-dimensional vascular structures within liver tissue.The resulting vascularized liver tissue model exhibits enhanced biological functionality,opening new opportunities for biomedical applications.展开更多
Immunoglobulin E(IgE)is a critical biomarker for diagnosing allergic and other immune diseases.However,traditional IgE detection methods are often time-consuming and require specialized operation.As an important bioma...Immunoglobulin E(IgE)is a critical biomarker for diagnosing allergic and other immune diseases.However,traditional IgE detection methods are often time-consuming and require specialized operation.As an important biomarker,researchers have used different strategies and explored various materials for designing IgE biosensors with highly sensitivity,specificity and cost-effectiveness.In this review,we comprehensively summarize recent advances in biosensing technologies for IgE detection,categorizing them into optical,electrochemical,and piezoelectric biosensors.It elaborates on the design principles,performance metrics,and comparative advantages of various platforms,such as surface plasmon resonance(SPR),fluorescence,electrochemical impedance spectroscopy(EIS),and quartz crystal microbalance(QCM).The integration of novel materials like aptamers and nanomaterials has significantly enhanced the performance of these biosensors.Biosensors present a promising alternative for rapid,sensitive,and efficient IgE detection,holding great potential for future clinical diagnostics and point-of-care testing applications.Future perspectives should highlight the discovery of novel materials,the application of artificial intelligence for aptamer development,and the synergy of optoelectronic sensing strategies to improve analytical performance for clinical diagnostics.展开更多
BACKGROUND: Acute lung injury(ALI) is a common and serious complication of severe acute pancreatitis(SAP). The study aimed to investigate the protective effect and mechanism of phosphatidylinositol-3 kinase(PI3K) inhi...BACKGROUND: Acute lung injury(ALI) is a common and serious complication of severe acute pancreatitis(SAP). The study aimed to investigate the protective effect and mechanism of phosphatidylinositol-3 kinase(PI3K) inhibitor Wortmannin in SAP associated with ALI.METHODS: Ninety rats were randomly divided into three groups: sham operation(SO) group(n=30), SAP group(n=30), and SAP+Wortmannin(SAP+W) group(n=30). SAP model was induced by retrograde injection of 4% sodium taurocholate into the biliopancreatic duct of rats. The rate of lung water content, myeloperoxidase(MPO), matrix metalloproteinase 9(MMP-9), protein kinase B(PKB), abdphosphorylation of protein kinase B(P-PKB) activity in the lung tissue were evaluated.RESULTS: In the SAP group, the p-PKB expression in the lung tissue began to rise at 3 hours after modeling, and peaked at 12 hours(P<0.05); the rate of lung water content, MPO and TNF-α activity were also gradually increased, and the degree of lung lesion gradually increased(P<0.05). In the SAP+Wortmannin group, the p-PKB expression in the lung tissue began to rise at 3 hours after modeling, and peaked at 12 hours; it was higher than that in the SO group(P<0.05), but significantly lower than that in the SAP group(P<0.05). The rest indicators in the SAP+Wortmannin group were also significantly decreased as compared with the SAP group(P<0.05).CONCLUSIONS: The expression of phosphatidylinositol-3 kinase/protein kinase B was elevated in severe pancreatitis rats with lung injury. This suggested that PI3 K signal transduction pathway is involved in the control and release of proinfl ammatory cytokines TNF-α, which may play an important role in the pathogenesis of severe acute pancreatitis associated with lung injury. This finding indicated that Wortmannin can block the PI3 K signal transduction pathway, and inhibit the release of infl ammatory factor TNF-α.展开更多
BACKGROUND:We investigated whether the use of a specially designed visual estimation tool may improve accuracy in quantifying blood volumes related to surface spreading.METHODS:A prospective,paired-control,single-blin...BACKGROUND:We investigated whether the use of a specially designed visual estimation tool may improve accuracy in quantifying blood volumes related to surface spreading.METHODS:A prospective,paired-control,single-blinded experimental study was performed at a medical university.Anesthesiologists and emergency medical personnel estimated various blood volumes on surfaces with varying absorptivity(carpet,towel,polyvinyl chloride,wooden fl ooring)in an experimental setting.We assessed the sensitivity of training blood volume quantifi cation using a self-designed visual estimation tool by comparing the accuracy of visual blood volume estimations before and after practical training with the tool.RESULTS:A total of 352 estimations by 44 participants were evaluated.Accurate estimations improved significantly from pre-training to post-training(P<0.05).The sensitivity of blood volume quantifi cation was 33.0%after training with the visual estimation tool.Estimations did not depend on age,profession,gender or years of the estimator’s professional experience.CONCLUSIONS:Training with a visual estimation tool by professional rescuers can improve the estimation accuracy of blood volumes spread on surfaces with varying absorptivity.展开更多
Objective：To evaluate the effectiveness and safety of the mobilization of peripheral blood hematopoietic stem cells by combining docetaxel with granulocyte colony-stimulating factor（G-CSF） in breast cancer patients...Objective：To evaluate the effectiveness and safety of the mobilization of peripheral blood hematopoietic stem cells by combining docetaxel with granulocyte colony-stimulating factor（G-CSF） in breast cancer patients.Methods：A total of 57 breast cancer patients were treated with docetaxel 120 mg/m2.When the white blood cell（WBC） count decreased to 1.0×109/L,patients were given G-CSF 5-g/kg daily by subcutaneous injection until the end of apheresis.Peripheral blood mononuclear cells（MNC） were isolated by Cobe Spectra Apheresis System.The percentage of CD34＋ cell was assayed by flow cytometry.Results：At a median 6 of days（range 3-8） after the administration of docetaxel,the median WBC count decreased to 1.08×109/L（range 0.20-2.31）.The median duration of G-CSF mobilization was 3 days（range 2-7）.The MNC collection was conducted 8-12 days（median 10 days） after docetaxel treatment.The median MNC was 5.35×108/kg（range 0.59-14.07）,the median CD34＋ cell count was 2.43×106/kg（range 0.16-16.69）.The CD34＋ cell count was higher than 1.00×106/kg in 47 of 57 cases（82.46%） and higher than 2.00×106/kg in 36 cases（63.16%）.The CD34＋ cell count was higher than 2.00×106/kg in 27 collections（23.68%）.The MNC count and the CD34＋ cell count were correlated with the bottom of WBC after docetaxel chemotherapy（r=0.364,0.502,P=0.005,0.000）.The CD34＋ cell count was correlated with the MNC count（r=0.597,P=0.000）.The mobilization and apheresis were well tolerated in all patients.Mild perioral numbness and numbness of hand or feet were observed in 3 cases.No serious adverse events were reported.Conclusion：Mobilization of peripheral blood hematopoietic stem cell by combining docetaxel with G-CSF was effective and safety in breast cancer patients.展开更多
The effect of the radial density configuration in terms of width, edge gradient and volume gradient on the wave field and energy flow in an axially uniform helicon plasma is studied in detail. A three-parameter functi...The effect of the radial density configuration in terms of width, edge gradient and volume gradient on the wave field and energy flow in an axially uniform helicon plasma is studied in detail. A three-parameter function is employed to describe the density, covering uniform,parabolic, linear and Gaussian profiles. It finds that the fraction of power deposition near the plasma edge increases with density width and edge gradient, and decays in exponential and ＂bumpon-tail＂ profiles, respectively, away from the surface. The existence of a positive second-order derivative in the volume density configuration promotes the power deposition near the plasma core, which to our best knowledge has not been pointed out before. The transverse structures of wave field and current density remain almost the same during the variation of density width and gradient, confirming the robustness of the m=1 mode observed previously. However, the structure of the electric wave field changes significantly from a uniform density configuration, for which the coupling between the Trivelpiece-Gould（TG） mode and the helicon mode is very strong, to non-uniform ones. The energy flow in the cross section of helicon plasma is presented for the first time, and behaves sensitive to the density width and edge gradient but insensitive to the volume gradient. Interestingly, the radial distribution of power deposition resembles the radial profile of the axial component of current density, suggesting the control of the power deposition profile in the experiment by particularly designing the antenna geometry to excite a required axial current distribution.展开更多
Hepatitis B virus(HBV)genotype B and C are two major genotypes that are prevalent in Asia and differ in natural history and disease progression.The impact of HBV genotypes on viral replication and protein expression h...Hepatitis B virus(HBV)genotype B and C are two major genotypes that are prevalent in Asia and differ in natural history and disease progression.The impact of HBV genotypes on viral replication and protein expression has been explored by the transfection of hepatoma cells with replication-competent HBV DNA,which mimics the later stages of the viral life cycle.However,the influence of HBV genotypes on the early events of viral infection remains undetermined,mainly due to the difficulties in obtaining sufficient infectious viral particles for infection assays.Here,we report that a high-titer HBV inoculum can be generated from the transient transfection-based cell model after optimizing transfection conditions and modifying the HBV-expressing construct.By performing in vitro infection assays using transiently transfected derived viruses,we found that clinical genotype C isolates possessed higher infectivity than genotype B isolates.Moreover,we identified a naturally occurring mutation sL21S in small hepatitis B surface protein,which markedly decreased the infectivity of HBV genotype C isolates,but not that of genotype B isolates.In summary,using infectious viral particles provided by the optimized transient transfection-based cell model,we have been able to investigate a wide range of HBV variants on viral infectivity,which may contribute to our understanding of the reasons for different clinical outcomes in HBV infections and the development of therapeutic drugs targeting the early stages of HBV life cycle.展开更多
Objective:To investigate the impact of the fliD gene on the biofilm formation of Helicobacter pylori(H.pylori).Methods:H.pylori fliD mutant was constructed using inverse PCR mutagenesis.The mobility of the bacteria an...Objective:To investigate the impact of the fliD gene on the biofilm formation of Helicobacter pylori(H.pylori).Methods:H.pylori fliD mutant was constructed using inverse PCR mutagenesis.The mobility of the bacteria and its adhesion ability to human epithelial cells were assessed using a motility assay and a fluorescein isothiocyanate staining adhesion assay,respectively.The formation of biofilm was evaluated using a pellicle assay and a crystal violet staining assay.The cy to-architecture of the biofilm was documented with scanning electron microscopy.Results:It was found that there was no significant difference in the levels of bacterial adhesion and the biofilm formation between the wild-type ATCC 43504 and the fliD mutant.Apart from a poor motility,the fliD mutant had a slightly delayed formation of its biofilm and an incomplete cy to-architecture of its biofilm.The bacterial cells residing in the biofilm of the fliD mutant showed a loose accumulation with less apparent crosslinking fibrils.Most of the mutant cells had truncated flagella.Conclusions:This study provides the preliminary evidences that fliD potentially regulates biofilm formation and is required for the motility of H.pylori.Further studies need to be performed in order to develop fliD as a novel target for vaccine or antimicrobial agent in future.展开更多
Abstract Objective We identify ionizing radiation-induced mitochondrial DNA （mtDNA） deletions in human lymphocytes and their distribution in normal populations. Methods Long-range polymerase chain reactions （PCR） ...Abstract Objective We identify ionizing radiation-induced mitochondrial DNA （mtDNA） deletions in human lymphocytes and their distribution in normal populations. Methods Long-range polymerase chain reactions （PCR） using two pairs of primers specific for the human mitochondrial genome were used to analyze the lymphoblastoid cell line following exposure to 10 Gy 6~Co y-rays. Limited-condition PCR, cloning and sequencing techniques were applied to verify the mtDNA deletions detected with long-range PCR. Human peripheral blood samples were irradiated with 0, 2 and 6 Gy ^60Co y-rays, and real-time PCR analysis was performed to validate the mtDNA deletions. In order to know the distribution of mtDNA deletions in normal population, 222 healthy Chinese adults were also investigated. Results Two mtDNA deletions, a 7455-bp deletion （nt475-nt7929 in heavy strand） and a 9225-bp deletion （nt7714 -nt369 in heavy strand）, occurring between two 8-bp direct repeats, were identified in lymphoblastoid cells using long-range PCR, limited-condition PCR and sequencing. These results were also observed for ^60Co y-rays irradiated human peripheral blood cells. Conclusion Two novel mtDNA deletions, a 7455-bp deletion and a 9225-bp deletion, were induced by ionizing radiation. The rate of the mtDNA deletions within a normal population was related to the donors＇ age, but was independent of gender.展开更多
Objective To investigate the therapeutic potential of tumor necrosis factor （TNF）-related apoptosis-inducing ligand （TRAIL）, a member of the TNF superfamily, and to analyze TRAIL-induced apoptosis in Jurkat cells....Objective To investigate the therapeutic potential of tumor necrosis factor （TNF）-related apoptosis-inducing ligand （TRAIL）, a member of the TNF superfamily, and to analyze TRAIL-induced apoptosis in Jurkat cells. Methods Expression of TRAIL receptors （DR4 and DR5） was detected by reverse transcriptase-polymerase chain reaction （RT-PCR）. Cytotoxic effects were determined by colony formation assay and a cell counting kit. The effects of recombinant TRAIL on apoptosis of Jurkat cells were determined by DNA fragmentation （DNA ladder） and PI staining. Changes in mitochondrial membrane potential were detected with JC-1 fluorescence. Results TRAIL inhibited the proliferation and induced intemucleosomal DNA fragmentation （characteristic of apoptosis） and loss of mitochondrial membrane potential. Conclusion Recombinant soluble TRAIL can be used as a therapy for cancer.展开更多
BACKGROUND In a previous paper,we reported a high prevalence of donor-specific antibody(DSA)in pediatric patients with chronic rejection and expressed the need for confirmation of these findings in a larger cohort.AIM...BACKGROUND In a previous paper,we reported a high prevalence of donor-specific antibody(DSA)in pediatric patients with chronic rejection and expressed the need for confirmation of these findings in a larger cohort.AIM To clarify the importance of DSAs on long-term graft survival in a larger cohort of pediatric patients.METHODS We performed a retrospective analysis of 123 pediatric liver transplantation(LT)recipients who participated in yearly follow-ups including Luminex testing for DSA at our center.The cohort was split into two groups according to the DSA status(DSA-positive n=54,DSA-negative n=69).Groups were compared with regard to liver function,biopsy findings,graft survival,need for re-LT and immunosuppressive medication.RESULTS DSA-positive pediatric patients showed a higher prevalence of chronic rejection(P=0.01),fibrosis(P<0.001)and re-transplantation(P=0.018)than DSA-negative patients.Class II DSAs particularly influenced graft survival.Alleles DQ2,DQ7,DQ8 and DQ9 might serve as indicators for the risk of chronic rejection and/or allograft fibrosis.Mean fluorescence intensity levels and DSA number did not impact graft survival.Previous episodes of chronic rejection might lead to DSA development.CONCLUSION DSA prevalence significantly affected long-term liver allograft performance and liver allograft survival in our cohort of pediatric LT.Screening for class II DSAs in combination with assessment of protocol liver biopsies for chronic antibodymediated rejection improved early identification of patients at risk of graft loss.展开更多
A novel kind of multi-core magnetic composite particles, the surfaces of which were respectively mo- dified with goat-anti-mouse IgG and antitransferrin receptor（anti-CD71）, was prepared. The fetal nucleated red blo...A novel kind of multi-core magnetic composite particles, the surfaces of which were respectively mo- dified with goat-anti-mouse IgG and antitransferrin receptor（anti-CD71）, was prepared. The fetal nucleated red blood cells（FNRBCs） in the peripheral blood of a gravida were rapidly and effectively enriched and separated by the mo- dified multi-core magnetic composite particles in an external magnetic field. The obtained FNRBCs were used for the identification of the fetal sex by means of fluorescence in situ hybridization（FISH） technique. The results demonstrate that the multi-core magnetic composite particles meet the requirements for the enrichment and speration of FNRBCs with a low concentration and the accuracy of detetion for the diagnosis of fetal sex reached to 95%. Moreover, the obtained FNRBCs were applied to the non-invasive diagnosis of Down syndrome and chromosome 3p21 was de- tected. The above facts indicate that the novel multi-core magnetic composite particles-based method is simple, relia- ble and cost-effective and has opened up vast vistas for the potential application in clinic non-invasive prenatal diag- nosis.展开更多
Human methylenetetrahydrofolate reductase(MTHFR),an essential enzyme in folate metabolism,catalyzes the conversion of5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate.Several polymorphisms in the MTHFR gene h...Human methylenetetrahydrofolate reductase(MTHFR),an essential enzyme in folate metabolism,catalyzes the conversion of5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate.Several polymorphisms in the MTHFR gene have been found to be associated with reduced enzyme activity and related to folic acid intake in relevant populations[1].展开更多
Metabolic liver diseases(MLD)are the second most common indication for liver transplantation(LT)in children.This is based on the fact that the majority of enzymes involved in various metabolic pathways are present wit...Metabolic liver diseases(MLD)are the second most common indication for liver transplantation(LT)in children.This is based on the fact that the majority of enzymes involved in various metabolic pathways are present within the liver and LT can cure or at least control the disease manifestation.LT is also performed in metabolic disorders for end-stage liver disease,its sequelae including hepatocellular cancer.It is also performed for preventing metabolic crisis’,arresting progression of neurological dysfunction with a potential to reverse symptoms in some cases and for preventing damage to end organs like kidneys as in the case of primary hyperoxalosis and methyl malonic acidemia.Pathological findings in explant liver with patients with metabolic disease include unremarkable liver to steatosis,cholestasis,inflammation,variable amount of fibrosis,and cirrhosis.The outcome of LT in metabolic disorders is excellent except for patients with mitochondrial disorders where significant extrahepatic involvement leads to poor outcomes and hence considered a contraindication for LT.A major advantage of LT is that in the post-operative period most patients can discontinue the special formula which they were having prior to the transplant and this increases their well-being and improves growth parameters.Auxiliary partial orthotopic LT has been described for patients with noncirrhotic MLD where a segmental graft is implanted in an orthotopic position after partial resection of the native liver.The retained native liver can be the potential target for future gene therapy when it becomes a clinical reality.展开更多
文摘Sonoclot analyzer has been widely used in many countries. But the reference intervals provided by the manufacturer were derived from only 45 participants, and there was no cut-off value for transfusion for Sonoclot analysis. This study aimed to establish reference intervals and transfusion criterion for Sonoclot analysis. Volunteers were recruited from healthy Chinese adults and patients undergoing cardiac surgery. Blood samples were withdrawn from forearm vein and measured for activated clotting time（ACT）, clot rate（CR）, platelet function（PF）, activated partial thromboplastin time（APTT）, fibrinogen concentration（FIB）, and platelet count（PLT）. The reference intervals were determined by the nonparametric method. Cut-off values were determined by the receiver operating characteristics curve. A total of 135 healthy volunteers and 281 patients were enrolled. The 95% reference intervals were 96–195 s, 22–51 signal U/min, 〉1.6 for ACT, CR, PF respectively. In the 281 patients, the results of APTT, FIB, PLT, ACT, CR, and PF ranged from 20.5–300.0 s, 0.28–4.11 g/L,（19.0–387.3）×109/L, 80–514 s, 2.9–74 signal U/min, and 0.1–5.1 respectively. The cut-off values for transfusion were 〉208, ≤14, and ≤1.3 for ACT, CR, PF respectively. The cut-off values of Sonoclot analysis were within the manufacturer＇s reference intervals, while they were outside the reference intervals established in this study. The results suggested that the manufacturer＇s reference intervals were not suitable for Chinese. The reference intervals and cut-off values established in this study will be helpful to Chinese patients.
文摘Purposes: Hypofibrinogenemia is usually complicated with severe and massive upper gastrointestinal (GI) bleeding, especially in liver cirrhosis. In Japan, neither fibrinogen concentrate nor cryoprecipitate (CRYO) is available for cases of acquired hypofibrinogenemia to recover the hemostatic level of fibrinogen rapidly. We prepared and produced in-house CRYO from fresh frozen plasma, and compared mortality between pre-implementation and post-implementation of CRYO. Methods: We performed a retrospective cohort study of patients admitted to our single tertiary medical center with upper GI bleeding from January 2011 to December 2016. The observational control period was from January 2011 to February 2013. From March 2013 to December 2016, we implemented the transfusion of CRYO, which was prepared and produced in our hospital. Clinical outcomes were compared between the patients in the two periods. Patients: Eleven patients in the control period and 10 in the intervention period were eligible for analysis. Results: In-hospital mortality (55% vs. 20%, P = 0.238) and mortality within 24 hour after admission (27% vs. 0%, P = 0.246) tended to be lower in the intervention period than in the control period, although the patients had more severe coagulation on admission than those in the control period. The plasma fibrinogen level before the treatment of hemostasis in the intervention period was higher than that in the control period (80 ± 9 mg/dL vs. 127 ± 15 mg/dL, P Conclusion: Implementation of in-house CRYO transfusion may reduce the rate of mortality due to severe upper GI bleeding.
文摘<strong>Objective:</strong> In Japan, red blood cell (RBC) solution is usually transported by car from a medical institution to the patient’s house for home transfusion. However, there are no regulations for transporting blood by car in the medical setting. Therefore, we assessed and compared the methods (containers) used for transporting the RBC solution by car. <strong>Materials and Methods:</strong> Irradiated RBC solution samples (280 mL) supplied by the Japan Red Cross Society were each divided into two bags. The quality of blood transported by car (1 - 2 hours) in an active transport refrigerator (ATR) (control group) was compared with that transported in a cooler, or styrofoam box (study group). We tested the hemolytic effects of transportation by car, storage, and filtration through a transfusion set on the lactate dehydrogenase (LD) levels in the RBC solutions. <strong>Results:</strong> Post-filtered LD levels were significantly higher in the RBC solutions transported in a cooler-box with inadequate temperature control when compared to those transported in an ATR with optimal temperature control. However, under conditions of optimal temperature control, the post-filtered LD levels were comparable in the control and study (both cooler and styrofoam boxes) group RBC solutions. <strong>Conclusion:</strong> Temperature management is critical for the maintenance of the quality of the RBC solution transported by car.
文摘Sickle cell disease is an autosomal recessive genetic disease. Vaso occlusive crisis (VOC) is frequently seen in such patients. Painful VOC is usually recurrent, of variable severity due to many factors and its management poses important challenge in the clinical practice. Few patients do not respond to standard therapies and continue to suffer severe pain for prolonged period or land to serious life threatening situation. The red cell exchange by aphaeresis is presumed to be one efficient alternative in this situation which can reduce the level of HbS below 40% - 50%. However, it is costly and not available everywhere. Both circumstances are common in our state where incidence of sickle cell disease is quite high. In such situations simple red cell exchange i.e. removing 1 unit (350 ml) of blood manually (by phlebotomy) and replacement with one unit normal red cell is effective. All of our four cases of SCA with severe acute VOC, are not responding to standard therapy but responded efficiently to this simple red cell exchange transfusion. Our present observation may pave the way of one simple, affordable, and effective measure to reduce the pain of severe acute VOC not responding to standard therapy. Moderate reduction of HbS by 8% - 14% by simple red cell exchange transfusion was associated with relief of pain of acute VOC;a new observation was reported in all our 4 cases which need to be validated by larger controlled studies.
文摘BACKGROUND Critically ill patients often present on admission or develop acute respiratory failure requiring intubation and application of positive pressure ventilation during their hospital stay.AIM To investigate and identify the epidemiological data,parameters associated with respiratory settings or the mechanics,and values related to arterial blood gases(ABGs)that are associated with outcomes in critically ill patients.METHODS A retrospective analysis of 131 patients[mean age,67.3 years;mean acute physiology and chronic health evaluation(APACHE)score,21.4]with acute respiratory failure requiring invasive mechanical ventilation was performed.The parameters that were statistically analyzed included demographic data,the presence of comorbidities,the presence of coronavirus disease 19(COVID-19),the respiratory rate(RR),peak airway pressure(Ppeak),minute ventilation(MV),positive endexpiratory pressure,and the values related to ABGs.In order to facilitate the statistical analysis,patients were evaluated and compared in groups:Survivors(n=41)vs non-survivors(n=90)and patients without acute kidney injury(AKI)(n=60)vs patients with AKI(n=71).Four endpoints were studied:Mortality,length of stay,duration of mechanical ventilation,and AKI.Group comparisons were performed using the following statistical tests:Theχ^(2) test with Yates’correction,Fisher’s exact test,the Mann-Whitney U test,and Spearman’s rank correlation analysis.Binary logistic regression analysis conducted after the univariate statistical tests facilitated the investigation of the independent predictors of mortality and AKI.A two-sided P value of less than 0.05 was considered the threshold of statistical significance.RESULTS Non-survivors presented statistically significant differences in terms of being older in age,the presence of comorbidities,elevated APACHE score,medical(vs surgical)reasons for admission,presence of COVID-19,lower pH at ABGs,lower values of the oxygenation ratio(arterial oxygen partial pressure to the fraction of inspired oxygen)and arterial oxygen partial pressure,and elevated values of Ppeak,positive end-expiratory pressure,RR,arterial carbon dioxide partial pressure,and MV.The factors identified as independent predictors of mortality were the presence of comorbidities,APACHE score,COVID-19 status,arterial carbon dioxide partial pressure,Ppeak,RR,and MV.COVID-19 presence and elevated values of RR and Ppeak were positively correlated with the other three endpoints(length of stay,the duration of mechanical ventilation in survivors,and the occurrence of AKI in the entire study population)that were studied.The other parameters exhibited a variable(either positive/negative,or no)correlation to the four endpoints under investigation.CONCLUSION Among all investigated outcome measures,COVID-19,Ppeak,and RR were strongly associated with all the endpoints studied,suggesting that proper interventions involving the modifiable respiratory parameters Ppeak and RR could improve the overall outcome in these patients.A novel finding of this study was the relationship between RR and AKI,which is worthy of further investigation.Future studies may explore the clinical interpretation of these findings to improve outcomes in critically ill patients with acute respiratory failure.
文摘BACKGROUND Low back pain is a major cause of disability worldwide,with intervertebral disc degeneration contributing to nearly 40%of cases.Conventional treatments focus on symptom relief without addressing the underlying degeneration.Platelet-rich plasma(PRP),a regenerative therapy rich in growth factors,offers potential therapeutic benefits through growth factor-mediated mechanisms,though clinical evidence is limited.AIM To evaluate the efficacy of intradiscal autologous PRP injection in reducing pain and improving function in patients with chronic lumbar disc prolapse.METHODS This pilot quasi-experimental study was conducted in tertiary care centre between July 2022 and June 2024.The study involved comparing the outcomes between group A(n=17)who failed to respond to conservative treatment measures and received intradiscal PRP injection with group B(n=22)who responded to conservative treatment.Visual Analogue Scale(VAS)and Functional Rating Index(FRI)scores were recorded at baseline,3 weeks,and 6 weeks for both the groups.RESULTS Forty patients were enrolled in the study.The PRP group demonstrated significant improvement in VAS and FRI scores compared to baseline.While both groups improved from their respective baselines,direct between-group comparisons are limited by baseline differences in symptom severity.Patients who failed conservative trial showed significant improvement following PRP intervention,with outcomes approaching those observed in physiotherapy responders.CONCLUSION Intradiscal PRP injection significantly improved pain and function in patients with lumbar disc disease,with clinical improvements that approached the level observed in physiotherapy responders,despite baseline differences in symptom severity.PRP shows promise as an effective treatment for lumbar disc pathology;however,these preliminary findings are limited by the small sample size and short follow-up,warranting larger trials with long-term evaluation.
基金the funding from the National Natural Science Foundation of China No.52275294the National Key Research and Development Program of China(No.2018YFA0703000)。
文摘Constructing an in vitro vascularized liver tissue model that closely simulates the human liver is crucial for promoting cell proliferation,mimicking physiological heterogeneous structures,and recreating the cellular microenvironment.However,the layer-by-layer printing method is significantly constrained by the rheological properties of the bioink,making it challenging to form complex three-dimensional vascular structures in low-viscosity soft materials.To overcome this limitation,we developed a cross-linkable biphasic embedding medium by mixing low-viscosity biomaterials with gelatin microgel.This medium possesses yield stress and self-healing properties,facilitating efficient and continuous three-dimensional shaping of sacrificial ink within it.By adjusting the printing speed,we controlled the filament diameter,achieving a range from 250μm to 1000μm,and ensuring precise control over ink deposition locations and filament shapes.Using the in situ endothelialization method,we constructed complex vascular structures and ensured close adhesion between hepatocytes and endothelial cells.In vitro experiments demonstrated that the vascularized liver tissue model exhibited enhanced protein synthesis and metabolic function compared to mixed liver tissue.We also investigated the impact of varying vascular densities on liver tissue function.Transcriptome sequencing revealed that liver tissues with higher vascular density exhibited upregulated gene expression in metabolic and angiogenesis-related pathways.In summary,this method is adaptable to various materials,allowing the rheological properties of the supporting bath and the tissue's porosity to be modified using microgels,thus enabling precise regulation of the liver tissue microenvironment.Additionally,it facilitates the rapid construction of three-dimensional vascular structures within liver tissue.The resulting vascularized liver tissue model exhibits enhanced biological functionality,opening new opportunities for biomedical applications.
文摘Immunoglobulin E(IgE)is a critical biomarker for diagnosing allergic and other immune diseases.However,traditional IgE detection methods are often time-consuming and require specialized operation.As an important biomarker,researchers have used different strategies and explored various materials for designing IgE biosensors with highly sensitivity,specificity and cost-effectiveness.In this review,we comprehensively summarize recent advances in biosensing technologies for IgE detection,categorizing them into optical,electrochemical,and piezoelectric biosensors.It elaborates on the design principles,performance metrics,and comparative advantages of various platforms,such as surface plasmon resonance(SPR),fluorescence,electrochemical impedance spectroscopy(EIS),and quartz crystal microbalance(QCM).The integration of novel materials like aptamers and nanomaterials has significantly enhanced the performance of these biosensors.Biosensors present a promising alternative for rapid,sensitive,and efficient IgE detection,holding great potential for future clinical diagnostics and point-of-care testing applications.Future perspectives should highlight the discovery of novel materials,the application of artificial intelligence for aptamer development,and the synergy of optoelectronic sensing strategies to improve analytical performance for clinical diagnostics.
文摘BACKGROUND: Acute lung injury(ALI) is a common and serious complication of severe acute pancreatitis(SAP). The study aimed to investigate the protective effect and mechanism of phosphatidylinositol-3 kinase(PI3K) inhibitor Wortmannin in SAP associated with ALI.METHODS: Ninety rats were randomly divided into three groups: sham operation(SO) group(n=30), SAP group(n=30), and SAP+Wortmannin(SAP+W) group(n=30). SAP model was induced by retrograde injection of 4% sodium taurocholate into the biliopancreatic duct of rats. The rate of lung water content, myeloperoxidase(MPO), matrix metalloproteinase 9(MMP-9), protein kinase B(PKB), abdphosphorylation of protein kinase B(P-PKB) activity in the lung tissue were evaluated.RESULTS: In the SAP group, the p-PKB expression in the lung tissue began to rise at 3 hours after modeling, and peaked at 12 hours(P<0.05); the rate of lung water content, MPO and TNF-α activity were also gradually increased, and the degree of lung lesion gradually increased(P<0.05). In the SAP+Wortmannin group, the p-PKB expression in the lung tissue began to rise at 3 hours after modeling, and peaked at 12 hours; it was higher than that in the SO group(P<0.05), but significantly lower than that in the SAP group(P<0.05). The rest indicators in the SAP+Wortmannin group were also significantly decreased as compared with the SAP group(P<0.05).CONCLUSIONS: The expression of phosphatidylinositol-3 kinase/protein kinase B was elevated in severe pancreatitis rats with lung injury. This suggested that PI3 K signal transduction pathway is involved in the control and release of proinfl ammatory cytokines TNF-α, which may play an important role in the pathogenesis of severe acute pancreatitis associated with lung injury. This finding indicated that Wortmannin can block the PI3 K signal transduction pathway, and inhibit the release of infl ammatory factor TNF-α.
文摘BACKGROUND:We investigated whether the use of a specially designed visual estimation tool may improve accuracy in quantifying blood volumes related to surface spreading.METHODS:A prospective,paired-control,single-blinded experimental study was performed at a medical university.Anesthesiologists and emergency medical personnel estimated various blood volumes on surfaces with varying absorptivity(carpet,towel,polyvinyl chloride,wooden fl ooring)in an experimental setting.We assessed the sensitivity of training blood volume quantifi cation using a self-designed visual estimation tool by comparing the accuracy of visual blood volume estimations before and after practical training with the tool.RESULTS:A total of 352 estimations by 44 participants were evaluated.Accurate estimations improved significantly from pre-training to post-training(P<0.05).The sensitivity of blood volume quantifi cation was 33.0%after training with the visual estimation tool.Estimations did not depend on age,profession,gender or years of the estimator’s professional experience.CONCLUSIONS:Training with a visual estimation tool by professional rescuers can improve the estimation accuracy of blood volumes spread on surfaces with varying absorptivity.
基金supported by a grant from the Beijing Capital Development Foundation for Medical Sciences (No. 2007-2053)
文摘Objective：To evaluate the effectiveness and safety of the mobilization of peripheral blood hematopoietic stem cells by combining docetaxel with granulocyte colony-stimulating factor（G-CSF） in breast cancer patients.Methods：A total of 57 breast cancer patients were treated with docetaxel 120 mg/m2.When the white blood cell（WBC） count decreased to 1.0×109/L,patients were given G-CSF 5-g/kg daily by subcutaneous injection until the end of apheresis.Peripheral blood mononuclear cells（MNC） were isolated by Cobe Spectra Apheresis System.The percentage of CD34＋ cell was assayed by flow cytometry.Results：At a median 6 of days（range 3-8） after the administration of docetaxel,the median WBC count decreased to 1.08×109/L（range 0.20-2.31）.The median duration of G-CSF mobilization was 3 days（range 2-7）.The MNC collection was conducted 8-12 days（median 10 days） after docetaxel treatment.The median MNC was 5.35×108/kg（range 0.59-14.07）,the median CD34＋ cell count was 2.43×106/kg（range 0.16-16.69）.The CD34＋ cell count was higher than 1.00×106/kg in 47 of 57 cases（82.46%） and higher than 2.00×106/kg in 36 cases（63.16%）.The CD34＋ cell count was higher than 2.00×106/kg in 27 collections（23.68%）.The MNC count and the CD34＋ cell count were correlated with the bottom of WBC after docetaxel chemotherapy（r=0.364,0.502,P=0.005,0.000）.The CD34＋ cell count was correlated with the MNC count（r=0.597,P=0.000）.The mobilization and apheresis were well tolerated in all patients.Mild perioral numbness and numbness of hand or feet were observed in 3 cases.No serious adverse events were reported.Conclusion：Mobilization of peripheral blood hematopoietic stem cell by combining docetaxel with G-CSF was effective and safety in breast cancer patients.
基金supported by National Natural Science Foundation of China(No.11405271)
文摘The effect of the radial density configuration in terms of width, edge gradient and volume gradient on the wave field and energy flow in an axially uniform helicon plasma is studied in detail. A three-parameter function is employed to describe the density, covering uniform,parabolic, linear and Gaussian profiles. It finds that the fraction of power deposition near the plasma edge increases with density width and edge gradient, and decays in exponential and ＂bumpon-tail＂ profiles, respectively, away from the surface. The existence of a positive second-order derivative in the volume density configuration promotes the power deposition near the plasma core, which to our best knowledge has not been pointed out before. The transverse structures of wave field and current density remain almost the same during the variation of density width and gradient, confirming the robustness of the m=1 mode observed previously. However, the structure of the electric wave field changes significantly from a uniform density configuration, for which the coupling between the Trivelpiece-Gould（TG） mode and the helicon mode is very strong, to non-uniform ones. The energy flow in the cross section of helicon plasma is presented for the first time, and behaves sensitive to the density width and edge gradient but insensitive to the volume gradient. Interestingly, the radial distribution of power deposition resembles the radial profile of the axial component of current density, suggesting the control of the power deposition profile in the experiment by particularly designing the antenna geometry to excite a required axial current distribution.
基金supported by grants of the National Youth Natural Science Foundation of China,China(82102380)the Youth Natural Science Foundation of Jiangsu Province(BK20200126)the National Youth Natural Science Foundation of China,China(81602400)。
文摘Hepatitis B virus(HBV)genotype B and C are two major genotypes that are prevalent in Asia and differ in natural history and disease progression.The impact of HBV genotypes on viral replication and protein expression has been explored by the transfection of hepatoma cells with replication-competent HBV DNA,which mimics the later stages of the viral life cycle.However,the influence of HBV genotypes on the early events of viral infection remains undetermined,mainly due to the difficulties in obtaining sufficient infectious viral particles for infection assays.Here,we report that a high-titer HBV inoculum can be generated from the transient transfection-based cell model after optimizing transfection conditions and modifying the HBV-expressing construct.By performing in vitro infection assays using transiently transfected derived viruses,we found that clinical genotype C isolates possessed higher infectivity than genotype B isolates.Moreover,we identified a naturally occurring mutation sL21S in small hepatitis B surface protein,which markedly decreased the infectivity of HBV genotype C isolates,but not that of genotype B isolates.In summary,using infectious viral particles provided by the optimized transient transfection-based cell model,we have been able to investigate a wide range of HBV variants on viral infectivity,which may contribute to our understanding of the reasons for different clinical outcomes in HBV infections and the development of therapeutic drugs targeting the early stages of HBV life cycle.
基金Supported by the Thailand Research Fund(MRG5580161)the Research Fund Project(ASHCU57002)+1 种基金the Faculty of Allied Health Sciences at Chulalongkorn Universitythe Graduate School Thesis Grant of Chulalongkorn University
文摘Objective:To investigate the impact of the fliD gene on the biofilm formation of Helicobacter pylori(H.pylori).Methods:H.pylori fliD mutant was constructed using inverse PCR mutagenesis.The mobility of the bacteria and its adhesion ability to human epithelial cells were assessed using a motility assay and a fluorescein isothiocyanate staining adhesion assay,respectively.The formation of biofilm was evaluated using a pellicle assay and a crystal violet staining assay.The cy to-architecture of the biofilm was documented with scanning electron microscopy.Results:It was found that there was no significant difference in the levels of bacterial adhesion and the biofilm formation between the wild-type ATCC 43504 and the fliD mutant.Apart from a poor motility,the fliD mutant had a slightly delayed formation of its biofilm and an incomplete cy to-architecture of its biofilm.The bacterial cells residing in the biofilm of the fliD mutant showed a loose accumulation with less apparent crosslinking fibrils.Most of the mutant cells had truncated flagella.Conclusions:This study provides the preliminary evidences that fliD potentially regulates biofilm formation and is required for the motility of H.pylori.Further studies need to be performed in order to develop fliD as a novel target for vaccine or antimicrobial agent in future.
基金supported by the National Natural Science Foundation of China(No.30570551and No.30870749)the Beijing Natural Science Foundation(No.7053073)
文摘Abstract Objective We identify ionizing radiation-induced mitochondrial DNA （mtDNA） deletions in human lymphocytes and their distribution in normal populations. Methods Long-range polymerase chain reactions （PCR） using two pairs of primers specific for the human mitochondrial genome were used to analyze the lymphoblastoid cell line following exposure to 10 Gy 6~Co y-rays. Limited-condition PCR, cloning and sequencing techniques were applied to verify the mtDNA deletions detected with long-range PCR. Human peripheral blood samples were irradiated with 0, 2 and 6 Gy ^60Co y-rays, and real-time PCR analysis was performed to validate the mtDNA deletions. In order to know the distribution of mtDNA deletions in normal population, 222 healthy Chinese adults were also investigated. Results Two mtDNA deletions, a 7455-bp deletion （nt475-nt7929 in heavy strand） and a 9225-bp deletion （nt7714 -nt369 in heavy strand）, occurring between two 8-bp direct repeats, were identified in lymphoblastoid cells using long-range PCR, limited-condition PCR and sequencing. These results were also observed for ^60Co y-rays irradiated human peripheral blood cells. Conclusion Two novel mtDNA deletions, a 7455-bp deletion and a 9225-bp deletion, were induced by ionizing radiation. The rate of the mtDNA deletions within a normal population was related to the donors＇ age, but was independent of gender.
基金This research was supported by Jiangsu Projects for Postdoctoral Research Funds (No. 0601025B) National Projects for Postdoctoral Research Funds (No. 20060390940).
文摘Objective To investigate the therapeutic potential of tumor necrosis factor （TNF）-related apoptosis-inducing ligand （TRAIL）, a member of the TNF superfamily, and to analyze TRAIL-induced apoptosis in Jurkat cells. Methods Expression of TRAIL receptors （DR4 and DR5） was detected by reverse transcriptase-polymerase chain reaction （RT-PCR）. Cytotoxic effects were determined by colony formation assay and a cell counting kit. The effects of recombinant TRAIL on apoptosis of Jurkat cells were determined by DNA fragmentation （DNA ladder） and PI staining. Changes in mitochondrial membrane potential were detected with JC-1 fluorescence. Results TRAIL inhibited the proliferation and induced intemucleosomal DNA fragmentation （characteristic of apoptosis） and loss of mitochondrial membrane potential. Conclusion Recombinant soluble TRAIL can be used as a therapy for cancer.
文摘BACKGROUND In a previous paper,we reported a high prevalence of donor-specific antibody(DSA)in pediatric patients with chronic rejection and expressed the need for confirmation of these findings in a larger cohort.AIM To clarify the importance of DSAs on long-term graft survival in a larger cohort of pediatric patients.METHODS We performed a retrospective analysis of 123 pediatric liver transplantation(LT)recipients who participated in yearly follow-ups including Luminex testing for DSA at our center.The cohort was split into two groups according to the DSA status(DSA-positive n=54,DSA-negative n=69).Groups were compared with regard to liver function,biopsy findings,graft survival,need for re-LT and immunosuppressive medication.RESULTS DSA-positive pediatric patients showed a higher prevalence of chronic rejection(P=0.01),fibrosis(P<0.001)and re-transplantation(P=0.018)than DSA-negative patients.Class II DSAs particularly influenced graft survival.Alleles DQ2,DQ7,DQ8 and DQ9 might serve as indicators for the risk of chronic rejection and/or allograft fibrosis.Mean fluorescence intensity levels and DSA number did not impact graft survival.Previous episodes of chronic rejection might lead to DSA development.CONCLUSION DSA prevalence significantly affected long-term liver allograft performance and liver allograft survival in our cohort of pediatric LT.Screening for class II DSAs in combination with assessment of protocol liver biopsies for chronic antibodymediated rejection improved early identification of patients at risk of graft loss.
文摘A novel kind of multi-core magnetic composite particles, the surfaces of which were respectively mo- dified with goat-anti-mouse IgG and antitransferrin receptor（anti-CD71）, was prepared. The fetal nucleated red blood cells（FNRBCs） in the peripheral blood of a gravida were rapidly and effectively enriched and separated by the mo- dified multi-core magnetic composite particles in an external magnetic field. The obtained FNRBCs were used for the identification of the fetal sex by means of fluorescence in situ hybridization（FISH） technique. The results demonstrate that the multi-core magnetic composite particles meet the requirements for the enrichment and speration of FNRBCs with a low concentration and the accuracy of detetion for the diagnosis of fetal sex reached to 95%. Moreover, the obtained FNRBCs were applied to the non-invasive diagnosis of Down syndrome and chromosome 3p21 was de- tected. The above facts indicate that the novel multi-core magnetic composite particles-based method is simple, relia- ble and cost-effective and has opened up vast vistas for the potential application in clinic non-invasive prenatal diag- nosis.
基金supported by A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)the Open-End Funds of the Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening[HY202105]。
文摘Human methylenetetrahydrofolate reductase(MTHFR),an essential enzyme in folate metabolism,catalyzes the conversion of5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate.Several polymorphisms in the MTHFR gene have been found to be associated with reduced enzyme activity and related to folic acid intake in relevant populations[1].
文摘Metabolic liver diseases(MLD)are the second most common indication for liver transplantation(LT)in children.This is based on the fact that the majority of enzymes involved in various metabolic pathways are present within the liver and LT can cure or at least control the disease manifestation.LT is also performed in metabolic disorders for end-stage liver disease,its sequelae including hepatocellular cancer.It is also performed for preventing metabolic crisis’,arresting progression of neurological dysfunction with a potential to reverse symptoms in some cases and for preventing damage to end organs like kidneys as in the case of primary hyperoxalosis and methyl malonic acidemia.Pathological findings in explant liver with patients with metabolic disease include unremarkable liver to steatosis,cholestasis,inflammation,variable amount of fibrosis,and cirrhosis.The outcome of LT in metabolic disorders is excellent except for patients with mitochondrial disorders where significant extrahepatic involvement leads to poor outcomes and hence considered a contraindication for LT.A major advantage of LT is that in the post-operative period most patients can discontinue the special formula which they were having prior to the transplant and this increases their well-being and improves growth parameters.Auxiliary partial orthotopic LT has been described for patients with noncirrhotic MLD where a segmental graft is implanted in an orthotopic position after partial resection of the native liver.The retained native liver can be the potential target for future gene therapy when it becomes a clinical reality.
