Objective Neuron purification is essential to procedure of various nerve cell experimental research, however, at present there is few reports on the effect of various factors on neural axons during purification. To find out a simple method of neuron purification, and to investigate the influence factors of corresponding purification culture in dorsal root gangl ion (DRG) tissue culture on β3-tubul in positive axon. Methods The DRGs were obtained from the 3 days neonatal SD rat microscopically and were made into cell suspension. Then, the amount of attached DRG neurons and non neuronal cells in poly-D-lysine (PDL) group, PDL/Laminin (PDL/LN) group and collagen-I (Col I) group was observed from 10 to 100 minutes. Then, the extension and arborization of β3-tubul in positive axons were observed after 72 hours completely randomised DRG tissue culture for the research of the influences among culture substrates (PDL, PDL/LN, and Col I), FBS (0, 5%, and 10%), 5 fluorouracil (5-Fu, 0, 20, and 40 μmol/L), and cytrarabine (Ara-C, 0, 10, and 20 μmol/L). Results Adherent cells were observed instantly after inoculation by inverted phase contrast microscope and inverted fluoresence microscope; after cell suspension was removed, adherent growth of DRGn cells and non-DRGn cells were still seen. In PDL group, the amount of NSE negative cells was significantly higher than that of NSE positive cells at 10 and 30 minutes (P lt; 0.05); the amount of NSE positive cells was significantly higher than that of NSE negative cells at 80, 90 and 100 minutes (P lt; 0.05). In PDL/LN gruop, there was no significant difference (P gt; 0.05) in the amount of NSE negative cells and NSE positive cells at 10, 20, 30, 40, and 50 minutes; the amount of NSE positive cells was significantly higher (P lt; 0.05) than that of NSE negative cells at 60, 70, 80, 90, 100 minutes. In Col I group, the amount of NSE negative cells was higher than that of NSE positive cells at 10-40 minutes, but showing no significant difference (P gt; 0.05); the amount of NSE positive cells was significantly higher (P lt; 0.05) than that of NSE negative cells at 70-100 minutes. At 72 hours after DRG tissue culture, the best result of β3-tubul in positive axon extension and arborization was obtained when the substrate level was PDL/LN, and the average length of PDL/LN level was significantly larger than that of other two substrates (P lt; 0.05). The highest number of β3-tubul in positive axon distal end was obtained at 5% concentration level of FBS (P lt; 0.05), but showing no significant differences in β3-tubul in positive axon length among three levels (P gt; 0.05). Both the most of β3-tubul in positive axon distal ends and the longest β3-tubul in positive axon average length were obtained at 0 μmol/L concentration level of 5-Fu, showing significant differences between 0 μmol/L level and 20, 40 μmol/L levels (P lt; 0.05). A similar result of β3-tubul in positive axon distal end was got at the 0 μmol/L level and 10 μmol/L level of Ara-C, which was significantly higher than that of 20 μmol/L level (Plt; 0.05). Conclusion? A purified DRG neuron suspension for neuron culture could be obtained via PDL differential attachment for 30 minutes. When DRG neuron culture, neuron special medium, PDL/LN substrate and 10 μmol/L Ara-C are recommended in β3-tubul in positive axon research.
Severe bee stings can trigger a systemic inflammatory response and multi-organ dysfunction, potentially resulting in fatality. Acute kidney injury (AKI) is a frequent complication in patients with severe bee stings, and conventional comprehensive treatment combined with various blood purification therapies is commonly employed in clinical practice to promptly manage the condition and reduce the average hospital stay duration. This article primarily delves into the significance of enhanced clinical nursing care for patients with bee stings-induced AKI undergoing blood purification therapy. Specifically, it underscores the importance of patient education regarding treatment-related considerations, nursing techniques for vascular access during treatment, potential complications, and corresponding nursing interventions.
Patients with severe acute kidney injury (AKI) often need renal replacement therapy (RRT）with a high morbidity and mortality. For patients with chronic renal failure, the aim of blood purification is renal replacement; but for patients with AKI, although customarily called RRT, the aim of blood purification is not “renal replacement”, but extracorporeal “renal support and protection”, that is, supporting and protecting temporally failed kidney, removing damage factors, avoiding renal reinjury and looking forward to restore renal function. This article provides a detailed explanation of the differences between renal replacement and renal support from the perspective of organ protection, as well as the key links of RRT and extracorporeal multiple organ support for patients with severe AKI.
With the deepening of current study and the innovation of perioperative management concept, there have been great advances in lung transplantation in recent years. The prognosis of patients has been significantly improved. At the same time, the role of various types of blood purification in the clinical monitoring and treatment of lung transplant patients is becoming increasingly prominent. This review aims to summarize the application and latest progress of in vitro blood purification such as renal replacement therapy, plasmapheresis and hemadsorption in the perioperative period of lung transplantation, and to provide a basis for further study.
bjective To separate the SO-Rb50 cells antigen corresponding to the monoclonal antibody of anti-retinoblastoma. Methods The antigen corresponding to the monoclonal antibody of anti-retinoblastoma was separated elementarily by ion-exchange chromatography, and was identified by dot-blotting using the monoclonal antibody of anti-retinoblastoma. The target protein band of the antigen was separated in light of sodium dodecyl sulfate-polyacrylamide gelelectrophoresis. Results A special unmixed band of SO-Rb50 cells antigen was separated with the relative molecular weight of 83×103.Conclusion The antigen corresponding to the monoclonal antibody of anti-retinoblastoma could be separated from SO-Rb50cells.(Chin J Ocul Fundus Dis，2003，19：152-155)
This article reviews Chinese nomenclature of renal replacement therapy and extracorporeal blood purification currently utilized to manage acute kidney injury and other organ dysfunction syndromes in critically ill patients, based on the recent reports of a consensus expert conference of Nomenclature Standardization Initiative Alliance. We provide a detailed description of the performance characteristics of membranes, filters, transmembrane transport of solutes and fluid, flows, and methods of measurement of delivered treatment, common definitions, components, techniques, and operations of the machines and platforms as well as the renal replacement therapy techniques in detail with the relevant technologies, procedures, operations, and recent developments in other extracorporeal therapies, including therapeutic plasma exchange, multiple organ support therapy, liver support, lung support, and blood purification in sepsis. We believe this nomenclature review will serve future use of terminology in publications, research, clinical operations and therapy platforms to enable consistent data collection and comparison.
To obtain recombinant human β2-microglobulin (rhβ2M) with properties of good solubility and high purity from E.coli, prokaryotic expression conditions were optimized and protein purification was performed in this study. After testing the effect of different IPTG concentrations, temperatures and induction times on the production of rhβ2M, the optimum expression conditions were determined, i.e. joining IPTG to final concentration being 0.8 mmol/L and inducing time 6 h and at temperature of 25℃. Under the optimum induction conditions, the ratio of soluble rhβ2M to soluble bacterial protein was 63.7%. After purified by Ni Sepharose 6 Fast Flow, the purity of rhβ2M achieved a greater value of 95%. Western blot analysis revealed that rhβ2M possessed the antigen property that specifically interacted with anti-β2M antibody.
Sepsis is a common clinical critical illness, which often leads to multiple organ damage including the kidney damage, which is difficult to treat and has a high mortality rate. In recent years, extracorporeal blood purification therapy has made some progress in the field of sepsis. There are a variety of blood purification modes to choose, but there is still no unified standard for the initiation timing of blood purification therapy. Clinicians mainly evaluate the indicators and the initiation timing of blood purification therapy according to the patient’s needs for renal function replacement and/or inflammatory mediator clearance. This article mainly summarizes and discusses the initiation timing of blood purification therapy in sepsis.
ObjectiveTo investigate the effect of pulsed colloid infusion combined with continuous blood purification (CBP) for treatment of severe capillary leak syndrome (CLS). MethodsAccording to random principle,61 patients were divided into a control group(n=21),a CBP1 group(n=18) and a CBP2 group(n=22). All patients of three groups received routine treatment according to international guidelines 2008 for management of severe sepsis. The patients in the control group also received pulsed infusion colloid combined lasix. The patients in the CBP1 and CBP2 groups also received continuous veno-venous hemofiltration(CVVH) for 72 hours. The patients in the CBP1 group received concentrated colloid infusion combined lasix,and the patients in the CBP2 group received concentrated colloid infusion combined removing fluid. Blood gas analysis and Impedance Cardiography was performed before and 24,48 and 72 hours after therapy. The angiopoietin-2(Ang-2) was measured. Also the length of ICU stay,duration of mechanical ventilation,and death rate of patients in 28 days were observed. ResultsCompared with the control group and the CBP1 group,the length of ICU stay(days) and duration of mechanical ventilation (days) in the CBP2 group were significantly shorter(P<0.05),and the death rate in 28 days was lower(P<0.05). The patients in the CBP2 group showed more reduction in the APACHEⅡ score compared with the CBP1 group after therapy(P<0.05). The oxygenation index in the CBP2 group respectively increased at 24,48 and 72 hours after therapy(P<0.05). Compared with the control group and the CBP1 group,the oxygenation index in the CBP2 group respectively increased at the same time(P<0.05). The thoracic fluid content (TFC) in the CBP2 group respectively decreased at 24,48 and 72 hours(P<0.05) after therapy,and decreased compared with the control group and the CBP1 group at the same time(P<0.05). The serum levels of Ang-2 in the CBP2 group respectively decreased at 24,48 and 72 hours after therapy(P<0.05),and decreased compared with the control group and the CBP1 group at the same time(P<0.05). ConclusionPulsed colloid infusion combined with continuous blood purification can reduce the severity of capillary leak and improves the outcome of patients with severe sepsis.
Objective To review the general approaches in isolation and purification of pancreatic islets and progress in several aspects. Methods The latest l iterature concerning acquisition of pancreatic islets was reviewed and analyzed interms of the choice of pancreatic islet donors, the digestion and isolation of pancreas, the purification of islet and the assay of outcome. Results The profile of the isolation and purification depends on the selection of reagents and methods of operation in every step and l inkup between every step. Conclusion Pancreatic islet transplantation is the most effective method to treat type 1 diabetes, the problem of inadequate sources of pancreatic islets could be resolved by the optimal process and the establ ishment of standardized operation.