Bead Method Research Articles

Quantitively Analyzing the Variation of Micrometer-Sized Microplastic during Water Treatment with the Flow Cytometry-Fluorescent Beads Method

Quantitively Analyzing the Variation of Micrometer-Sized Microplastic during Water Treatment with the Flow Cytometry-Fluorescent Beads Method

Application of magnetic bead method in detecting coagulation function.

Blood samples indicative of jaundice, lipemia, hemolysis, and others are often encountered in the laboratory, and such features impact greatly on the detection of coagulation items. To understand the anti-interference ability of the magnetic bead method automatic coagulation instrument and the optical method automatic coagulation instrument against jaundice, lipemia, and hemolysis, anti-interference experiments of prolonged prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), and fibrinogen (FIB) were conducted using ExC810 (magnetic bead method) and CS5100 (optical method). Interference samples were prepared with bilirubin, hemoglobin, and lipids, while control samples were prepared with NaOH and distilled water. The samples contained different values of PT, APTT, TT, and FIB and were detected by magnetic bead method and optical method, respectively. The relative deviation was calculated according to the formula. In the anti-interference experiment of PT, APTT, TT, and FIB (jaundice, lipemia, hemolysis), the deviation between the test results and the control results with the addition of interfering substances tested by ExC810 was lower overall than that of CS5100. However, after the addition of interfering substances, most of the items were not detected by CS5100 in the anti-lipidemia experiment. When testing coagulation function, the magnetic bead method has better anti-interference recognition of jaundice, lipemia, and hemolysis than the optical method, and its anti-lipemic interference has a particularly obvious advantage.

Preparing PAMAM-NK4 nano complexes and examining their in vitro growth suppression effects in breast cancer.

This study sought to examine the suppression of the NK4 (which is a fragment that originates from the trypsin digestion of the hepatocyte growth factor) gene as mediated by new nano material polyamidoamine (PAMAM) dendrimers in the growth of breast cancer cells MDA-MB-231 and MCF-7, and the therapeutic effects in a nude mice model of transplanted tumor cell MDA-MB-231. We built PAMAM-NK4 nano particles and detected the in vitro transfection rate. Nano complexes and blank plasmid PAMAM dendrimers were transfected to MDA-MB-231 and MCF-7 cells, respectively. The western-blotting method, MTT experiment method, and bead method were used to detect the effects of the nano complexes on NK4 protein expression, cell proliferation, and cell apoptosis. The nude mice model of transplanted tumor cell MDA-MB-231 comprised 40 nude female mice who were subject to injections. The mice were randomly divided into four groups, comprising 10 mice per group. The control, blank plasmid and treatment groups were subcutaneously injected with 0.2 mL of 0.9% NaCl (Sodium chloride) solution, 0.2 mL of plasmid solution (including 100 µg PAMAM pcDNA3.1(-) blank plasmid nano complexes) and 0.2 mL of plasmid solution (including PAMAM-NK4 100 µg) beside the tumor inoculation spot, respectively. The positive control group was intraperitoneally injected with 0.2 mL of doxorubicin solution, including 100 µg doxorubicin. Western blotting was used to detect the NK4 protein expression of the transplanted tumor tissues of the various groups. NK4 protein was successfully expressed in MDA-MB-231 and MCF-7 cells transfected with PAMAM-NK4 nano particles, and cell proliferation was suppressed and cell apoptosis was induced. The tumor volumes and masses of the treatment and positive control groups were obviously smaller than those of the control group. The differences were statistically significant (P<0.05). The treatment group had an obviously higher mean value of NK4 protein expression than the control group. The differences were statistically significant (P<0.05). PAMAM-NK4 nano complexes suppress the growth of the breast cancer cells MDA-MB-231 and MCF-7, and had a treatment effect on this tumor nude mice model of breast cancer cells.

Comparative study of two high-efficiency methods for purifying spermatogonial stem cells

Objective: To compare the advantages and disadvantages of the differential attachment method and immunomagnetic bead method for purification of mouse spermatogonial stem cells (mSSCs). Methods: Ten male C57BL/6 mice aged 12～15 days were selected and sacrificed by cervical dislocation. Testes were collected and the seminiferous tubule single cell suspension was obtained by enzymatic digestion. mSSCs were purified by using the differential attachment method and immunomagnetic bead method respectively. Then a detailed comparison of the two methods in terms of cell number, separation efficiency, and impact on cell proliferation and growth was conducted. Results: Both of the methods could isolate and purify stem cells from single cell suspension of mouse seminiferous tubules. mSSCs showed typical grape cluster-like clones in vitro culture, which could be continuously cultured and proliferated for over 3 months in vitro. The testes of 10 mice could obtain 3×105±0.4×105 mSSCs (n=5) by differential attachment method, cell recovery rate (the number of cells after purification/the number of cells of the single cell suspension of seminiferous tubules) was 1.5%±0.1%; 6×105±0.4×105 mSSCs (n= 5) could be obtained by immunomagnetic bead method. The recovery rate was about 3%±0.1%, and the number of stem cells obtained by the immunomagnetic bead method was higher. The stem cells obtained by the differential attachment method were more pure, because the stem cell colonies were preferentially obtained after 5 days of in vitro culture, while the stem cells obtained by the immunomagnetic bead method needed to be cultured for about 10 days before the obvious cell colonies could be observed, but the two types of purification method had no obvious effect on the long-term growth of cells in vitro. Conclusion: Both methods can get high quality mSSCs, but both methods have their own advantages and disadvantages. The differential attachment method is more economical and practical than the other, it does not require special equipment, but the stem cell number obtained is relatively lower and the time needed is longer.

Real-Time Recombinase Polymerase Amplification (RPA) Detection of Pseudomonas aeruginosa Using Magnetic Nano-Beads for DNA Extraction

Pseudomonas aeruginosa (P. aeruginosa) is a ubiquitous nonfermentative gram-negative bacillus, residing in nature widely as a conditional pathogen that is commonly isolated from nosocomial infection cases, having a larger genome (5.5–7 Mbp). P. aeruginosa possesses great environmental adaptability and higher mutation rates, which accounts for its ability to resist antibiotics. Furthermore, multi-antibiotics-resistant P. aeruginosa has recently been established to be responsible for increased nosocomial infection incidence. Therefore, to detect, diagnose, and treat this life-threatening infection early enough, we introduced a real-time recombinase polymerase amplification (RPA) method of rapid P. aeruginosa detection, which is time-saving, convenient, highly sensitive, and more specific than the traditional bacterial culture method. System effectiveness and efficiency were then determined by a series of experiments, where specific RPA primers and probes accurately distinguished P. aeruginosa from other common pathogenic bacteria with optimal RPA reaction procedures set at 39 °C for 20 min. Additionally, specific primers and probes were designed based on the oprF gene sequence of P. aeruginosa. Subsequently, a real-time RPA system was used to analyze 20 cases of different clinical samples for P. aeruginosa detection in blood, open wound swab, and sputum. Results indicated that this system was suitable for bacterial analysis of common clinical sample types. The DNA extraction method used in this study was based on the magnetic beads method. In conclusion, this study established that the real-time RPA system was a rapid, specific, and sensitive method for clinical P. aeruginosa detection, which can be integrated into automatic high-throughput detection machines.

Differential white spot syndrome virus-binding proteins in two hemocyte subpopulations of Chinese shrimp (Fenneropenaeus chinensis)

A number of white spot syndrome virus (WSSV)-binding proteins have been identified previously in the hemocytes of Fenneropenaeus chinensis. In order to further investigate the differential WSSV-binding proteins in hemocyte subpopulations, granular hemocytes and hyalinocytes were sorted from WSSV-infected shrimp by immunomagnetic bead (IMB) method. The results of ELISA and immuno-dot blot assay showed that the WSSV-binding activity of granular hemocytes proteins was much stronger than that of hyalinocytes proteins. And the percentage of WSSV-positive granular hemocytes was significantly higher than that of hyalinocytes post WSSV infection, indicating that granular hemocytes were more susceptible to WSSV infection. Moreover, a total of 9 WSSV-binding proteins were successfully identified in granular hemocytes and hyalinocytes by two-dimensional virus overlay protein binding assay (2D-VOPBA) and MALDI-TOF MS analysis, of which 3 binding proteins (arginine kinase, protease 1 and transglutaminase) existing in both hyalinocytes and granular hemocytes and 6 proteins (F1ATP synthase β-chain, hnRNPs, GAPDH, RACK1, β-actin and cellular retinoic acid) detected only in granular hemocytes. Among these identified WSSV-binding proteins, the transglutaminase (TG) was further recombinantly expressed, and the recombinant TG could be bound with WSSV. Subsequently, quantitative real-time PCR analysis showed that differential expression levels of WSSV-binding proteins were observed in granular hemocytes and hyalinocytes. The results of this study revealed that the WSSV-binding proteins were differentially expressed in granular hemocytes and hyalinocytes, which provided a deeper insight into the interaction between WSSV and hemocyte subpopulations.

Developing Two Rapid Protein Extraction Methods Using Focused-Ultrasonication and Zirconia-Silica Beads for Filamentous Fungi Identification by MALDI-TOF MS.

Filamentous fungi identification by Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been challenging due to the lack of simple and rapid protein extraction methods and insufficient species coverage in the database. In this study, we created two rapid protein extraction methods for filamentous fungi: a one-step zirconia-silica beads method (ZSB) and a focused-ultrasonication method (FUS). The identification accuracy of two methods were evaluated with the VITEK MS, as well as number of spectra peaks and signal-to-noise ratio (S/N) with M-Discover 100 MALDI-TOF MS compared to the routine method. The better method was applied to build a filamentous fungi in-house spectra library for the M-Discover 100 MS, and then another one and routine method were performed in parallel to verify the accuracy and commonality of the in-house library. Using the two optimized methods, the dedicated operating time before MALDI-TOF MS analysis was reduced from 30 min to 7 (ZSB) or 5 (FUS) min per sample, with only a few seconds added for each additional strain. And both two methods identified isolates from most mold types equal to or better than the routine method, and the total correct identification rate using VITEK MS was 79.67, 76.42, and 76.42%, respectively. On the other hand, the two rapid methods generally achieved higher maximum and minimum S/N ratios with these isolates tested as compared to the routine method. Besides, the ZSB method produced overall mean of maximum and minimum S/N ratio higher than that by FUS. An in-house library of M-Discover MS was successfully built from 135 isolates from 42 species belonging to 18 genera using the ZSB method. Analysis of 467 isolates resulted in 97.22% correctly identified isolates to the species level by the ZSB method versus 95.50% by the routine method. The two novel methods are time- and cost-effective and allow efficient identification of filamentous fungi while providing a simplified procedure to build an in-house library. Thus, more clinical laboratories may consider adopting MALDI-TOF MS for filamentous fungi identification in the future.

Functional Magnetite Nanoparticle: A Review on the Particles Lysis and Nucleic Acid Separation

The functional magnetite nanoparticles are one of the most important functional materials for nucleic acid separation. Cell lysis and magnetic separation are two essential steps involve in optimizing nucleic acid extraction using the magnetic beads method. Many coating materials, coupling agents, chemical cell lysis, and several methods have been proposed to produce the specific desired properties for nucleic acid extraction. The important properties, such as biocompatibility, stability, linking ability, hydrophobicity, and biodegradable, were considered. The appropriate coating material of magnetite core and coupling agent are necessary to give biomolecules a possibility to link with each other through chemical conjugation. In this review, progress in functional magnetite nanoparticles to optimize the high binding performance in nucleic acid extraction is discussed.

Study of the distribution of KIR3DL2 alleles among ethnic Han Chinese from Zhejiang

To study the distribution of KIR3DL2 alleles among ethnic Han Chinese from Zhejiang. Genomic DNA was extracted by using a magnetic bead method. The full sequence of the KIR3DL2 gene was amplified with four pairs by PCR primers. The coding regions of 208 unrelated ethnic Han Chinese blood donors were analyzed using a BigDye Terminator v3.1 Sequencing Kit. The genotypes were assigned based on the nucleotide polymorphism of the KIR3DL2 gene. Among the 208 samples, 133 were KIR3DL2 heterozygotes and 75 were homozygotes. Forty six KIR3DL2 genotypes were detected. Respectively, 70, 33 and 23 individuals were found to have a KIR3DL2*00201/KIR3DL2*00201, KIR3DL2*00201/KIR3DL2*00701, and KIR3DL2*00201/KIR3DL2*01001 genotype. Twenty-two KIR3DL2 alleles were discovered, and the frequencies of KIR3DL2*00201, KIR3DL2*00701 and KIR3DL2*01001 were 57.45%, 13.46% and 9.13%, respectively. The distribution of KIR3DL2 alleles among ethnic Han Chinese in Zhejiang has been determined and fits the criteria for genetic polymorphism.

Successful Urine Multiplex Bead Assay to Measure Lupus Nephritis Activity

IntroductionLupus nephritis (LN) confers a poor prognosis, mainly from lack of effective laboratory tests to diagnose and to evaluate therapies. We have previously shown that a set of 6 urinary biomarkers (NGAL, KIM-1, MCP-1, adiponectin, hemopexin, and ceruloplasmin) are highly sensitive and specific to identify adult and pediatric patients with active LN using renal biopsy as reference standard. Using these combinatorial urinary biomarkers, the Renal Activity Score for Lupus (RAIL) score was established, with biomarkers measured by enzyme-linked immunosorbent assay (ELISA). To enhance clinical utility of the biomarkers and RAIL, we tested the performance of RAIL with biomarkers measured by ELISA to that of biomarkers measured by the bead multiplex method, hypothesizing that the multiplex bead method would be comparable.MethodsSpot urine samples (n = 341) of 46 patients aged 20 to 73 years with or without LN were used. Samples were assayed both by ELISA and multiplex using LUMINEX. RAIL scores and biomarker quantities were assessed for agreement with intraassay correlation coefficients and compared using Bland−Altman and regression.ResultsBiomarker measurement by LUMINEX was successful for NGAL, KIM-1, MCP-1, and adiponectin, but not for ceruloplasmin and hemopexin. There was good agreement of the RAIL obtained from these 4 biomarkers, irrespective of assay method (intraclass correlation coefficient [ICC] = 0.78, 95% confidence interval [CI] = 0.78−0.82). The RAIL scores from 4 biomarkers further correlated with those when considering all 6 biomarkers (ICC = 0.97, 95% CI = 0.96−0.98).ConclusionThe LUMINEX platform allows for the convenient and simultaneous measurement of 4 RAIL biomarkers. RAIL scores considering only these 4 biomarkers may be sufficient to accurately capture LN activity.

Helicobacter pylori causes delayed gastric emptying by decreasing interstitial cells of Cajal.

Helicobacter pylori (HP) infection is one of the most frequent bacterial infections in humans and is associated with the pathogenesis of gastric motility disorders such as delayed gastric emptying (DGE). Although HP infection is considered to delay gastric emptying, there has been little research on the underlying mechanism. Gastric motility involves interactions among gastrointestinal hormones, smooth muscle, enteric and extrinsic autonomic nerves and interstitial cells of Cajal (ICCs), and ICCs play an important role in gastrointestinal motility. Mutation or loss of stem cell factor (SCF) expression is known to reduce the number of ICCs or alter the integrity of the ICC network, contributing to gastrointestinal dysmotility. The aim of the present study was to investigate whether a reduction in ICCs contributes to the DGE caused by HP. A mouse model of HP infection was established and gastric emptying was compared between HP-infected and uninfected mice using the bead method. In addition, ICC counts and SCF expression levels in gastric tissue were evaluated using immunohistochemistry and western blotting, respectively. The results revealed that gastric emptying was significantly slower, the number of ICCs in gastric tissue was significantly reduced and the protein level of SCF in gastric tissue was significantly decreased in HP-infected mice compared with uninfected mice. Therefore, it may be concluded that HP reduced the number of ICCs by decreasing the expression of SCF protein in gastric tissue, thereby causing DGE.

Evaluation of Bonding Gap Control Methods for an Epoxy Adhesive Joint of Carbon Fiber Tubes and Aluminum Alloy Inserts.

The aim of this paper is to compare two methods of epoxy adhesive bond gap control: one with a geometrical (mechanical) solution and the other with glass beads, which have the diameter of the desired bond gap and are mixed with an epoxy adhesive. The adhered materials were carbon fiber composite tubes and aluminum alloy inserts, which were used as wishbones in a suspension system of a motorsport vehicle. It was assumed that the gap thickness would be equal to 0.2 mm and the length of a bond would be 30 mm. The internal diameter of the tubes was 14 mm and 18 mm, whereas the inserts’ external diameter was 13.6 mm and 17.6 mm. Their surface has been subjected to mechanical treatment with sand paper starting from 240 grit up to 400. The adhesives used were EA 3425 and EA 9466 cured at 80 °C for 2 h. The results showed that the glass beads method provides more consistent and better results as compared to the geometrical (mechanical) method. Further study in the area of fatigue and interfacial failure modes could be useful.

Apatinib Inhibits Stem Properties and Malignant Biological Behaviors of Breast Cancer Stem Cells by Blocking Wnt/β-catenin Signal Pathway through Downregulating LncRNA ROR

Cancer stem cells could influence tumor recurrence and metastasis. To develop a new effective treatment modality targeting breast cancer stem cells (BCSCs) and to explore the role of Apatinib in BCSCs. BCSCs were isolated from MDA-MB-231 cells by the immune magnetic beads method. BCSCs were treated with Apatinib, lentiviral plasmids (lncRNA ROR), and iCRT-3 (Wnt pathway inhibitors). Viability, colony numbers, sphere numbers, apoptosis, migration, invasion of BCSCs were detected by MTT, colony formation, tumorsphere, flow cytometry, wound-healing, transwell assays, respectively. The expressions of markers (ABCG2, CD44, CD90, and CD24), epithelial-mesenchymal transition (EMT)-related molecules (Ecadherin, N-cadherin, Vimentin, MMP-2, MMP-9), and Wnt/β-catenin pathway-related proteins (Wnt3a, Wnt5a, β-catenin) in breast cancer stem cells were determined by performing Western blot and qRT-PCR analysis. Apatinib decreased the viability and colony numbers of BCSCs in a concentration-dependent manner, and it also reduced sphere numbers, suppressed migration, invasion and lncRNA ROR expression, and induced apoptosis of BCSCs. However, these results were partially reversed by lncRNA ROR overexpression. Apatinib suppressed stem property, EMT process, and Wnt/β-catenin pathway in BCSCs, which was partially reversed by lncRNA ROR overexpression. Moreover, lncRNA ROR overexpression increased the colony and sphere numbers and promoted the cell viability, apoptosis inhibition, migration, and invasion of BCSCs, but these effects were partially reversed by iCRT-3. LncRNA ROR overexpression increased the stem property, EMT process, and Wnt/β-catenin pathway, which were partially counteracted by iCRT-3. Apatinib inhibited stem property and malignant biological behaviors of BCSCs by blocking the Wnt/β-catenin signal pathway through down-regulating lncRNA ROR.

Design of Detection Module in Coagulometers Based on Dual-magnetic Circuit Beads Method

Coagulometer, known as blood coagulation analyzer, is a product that can provide accurate test results for medical diagnosis and treatment analysis by detecting a series of items closely related to thrombosis and hemostasis in coagulation reaction. On the basis of previous traditional methods, and with our deep understanding about the principles of hemagglutination detection, we propose a hemagglutination detection method by using the dual-magnetic circuit beads method. Then, the corresponding hemagglutination detection module is designed. The coagulation time of plasma can be measured by detecting the movement of the magnetic beads when the magnetic field intensity is appropriate. The activated partial thromboplastin time(APTT) of plasma is tested when the most suitable magnetic field intensity is found. The results preliminarily show that this blood coagulation test method is valid and the corresponding test module has a potential value in business.

Promoter Hypermethylation of LATS1 Gene in Oral Squamous Cell Carcinoma (OSCC) among North Indian Population.

Background: LATS1 (Large Tumor Suppressor, isoform 1) is a gene that forms a complex with the cyclin-dependent kinase, CDK1, and regulates cell cycle progression. Genetic modifications lead to a loss in the activity of LATS1 gene. OSCC is the most commonly emerging cancer caused by genetic as well as epigenetic changes. Epigenetics changes vary from one population to another because these are influenced by dietary factors and environmental factors. Tobacco chewing and smoking has been reported as major risk factors in OSCC. No report was found in the previous literature showing promoter hypermethylation of LATS1 gene. Methods:A total of 50 OSCC patients and 20 normal individuals were recruited in this study. Blood samples (50) from OSCC patients and blood samples (20) from healthy individuals as controls were used in the present study. Isolation of genomic DNA was carried out from blood using the standard phenol-chloroform extraction. Further Isolated DNA was modified with sodium bisulfite using the agarose bead method and finally, the methylation studies of LATS1 gene were carried out using Methylation-Specific PCR (MSP-PCR). Results:19 out of 50 patients (38.0%) were found to be methylated for LATS1 gene.; a statistically significant result was obtained (p -value= < 0.05) with an odds ratio of 0.37 in cases compared to controls. The status of methylation of LATS1 genes was also found to be statistically significantly associated with smokers and tobacco chewers (p-value = < 0.05). The methylation of LATS1 gene showed a significant risk of developing OSCC in patients. Conclusion:These results suggest that the LATS1 gene may provide a better alternative as a diagnostic biomarker. This is the first report on the promoter hypermethylation of LATS1 gene in OSCC patients among the North Indian population.

Comparison of different DNA extraction procedures for Eimeria oocysts

A study was carried out to compare the efficacy of different DNA extraction procedures for Eimerian oocysts. A total of 753 litter and fecal samples and intestines were collected from desi poultry farms and slaughter houses in Bengaluru, Karnataka. Out of five different methods of DNA extraction procedures the glass bead method was found to be the best method. Since the above method involves breaking of the oocysts and sporocysts walls and dissolving the sporozoites membrane. This greatly simplifies the currently used DNA extraction procedures for Eimeria species. The liquid nitrogen method was also found to be good but was more critical and time consuming. Other methods such as sodium hypochlorite, direct and sonication has failed to yield DNA from Eimeria oocysts. The present study indicates that glass beads method yielded better quality of DNA for molecular analysis and it is suitable for both large and small number (˂1x103) of oocysts in a sample. Hence, can be used as a routine procedure for DNA extraction for Eimerian oocysts.

The Complex Relationship between C4b-Binding Protein, Warfarin, and Antiphospholipid Antibodies

Low levels of total C4b-binding protein (C4BPt), a circulating inhibitor of the classical/lectin complement pathways, were observed in patients with antiphospholipid antibodies (aPLs) and during warfarin treatment. To investigate the associations between aPL and C4BPt in patients with persistently positive (++) aPL, with/without clinical manifestations and systemic lupus erythematosus (SLE), and in controls. Furthermore, we explored the impact of anticoagulation on C4BPt and in relation to complement activation. In a cross-sectional design we investigated defined subgroups: primary (p) antiphospholipid syndrome (APS, N = 67), aPL++ individuals without clinical manifestations (aPL carriers, N = 15), SLE-aPL++ (N = 118, among them, secondary [s] APS, N = 56), aPL negative (-) SLE (SLE-aPL-, N = 291), and 322 controls. Clinical characteristics, including treatment, were tabulated. C4BPt was determined with a magnetic bead method. Complement proteins (C1q, C2, C3, C4, C3a, C3dg, sC5b-9, factor I [FI]) were measured. A mediation analysis was performed to decompose the total effect of aPL++ on C4BPt into the direct and indirect effects of aPL++ through warfarin. Overall, C4BPt is 20% decreased in aPL++ patients, regardless of SLE, APS, clinical manifestations, and aPL profile. C4BPt levels associate positively with complement proteins C1q, C2, C3, and C4, and negatively with complement activation product C3dg. In the SLE group, warfarin treatment contributes to approximately half of the C4BPt reduction (9%) CONCLUSION: Both aPLs and warfarin are associated with C4BPt reduction. Complement activation in aPL++ patients may partly be explained by impaired inhibition through depressed C4BPt levels. Further studies are needed to understand the clinical implications.

The Value of Combined Detection of D-dimer and CD62p in Judging the Severity of Acute Cerebral Infarction and Short-Term Prognosis.

Objective To explore the value of combined detection of peripheral blood P-selectin (CD62p) and D-dimer (D-dimer) in the judgment of acute cerebral infarction severity and short-term prognosis. Methods 268 patients with acute cerebral infarction from February 2015 to February 2019 were selected as the observation group. According to the National Institute of Health stroke scale, there were 90 cases (SCI group), 88 cases (MOCI group), and 90cases (MICI group) in the severe, moderate, and mild cerebral infarction groups, respectively. In the same period, 80 cases of healthy people served as the Normal group. Use flow cytometry to detect CD62p in peripheral blood and magnetic bead method to detect D-dimer level within 24 hours of onset. Logistic regression was used to analyze whether the two are factors affecting the short-term prognosis of acute cerebral infarction, and the ROC curve was drawn to evaluate the value of the combined detection of the two in the short-term prognosis of patients with acute cerebral infarction. Results Peripheral blood D-dimer and CD62p levels (2.95 ± 0.76 ng/l, 34.03 ± 5.29 ng/l) in the SCI group were higher than those in the MOCI group (2.30 ± 0.51 ng/l, 27.58 ± 5.56 ng/l) and the MICI group (1.87 ± 0.40 ng/l, 19.60 ± 3.98 ng/l); the difference between the groups was statistically significant (P < 0.05). Logistic regression analysis showed that D-dimer and CD62p were independent risk factors affecting the poor prognosis of patients with acute cerebral infarction (OR values were 3.752 and 1.213, and 95% CI were 1.612-7.934 and 1.093-1.342, respectively, both P < 0.05). The AUC of D-dimer combined with CD62p for predicting poor prognosis of acute cerebral infarction is 0.859, which is better than D-dimer and CD62p alone. Conclusion Peripheral blood D-dimer combined with CD62p detection is helpful for the risk stratification and short-term prognosis assessment of patients with acute cerebral infarction. Clinical detection is of great significance for the prevention and monitoring of disease development.

Application of Microfluidic Technology in Pharmaceutical Analysis

Microfluidic gadgets existing one-of-a-kind benefits for the development of effective medication provider bits, cell-free healthy protein synthesis systems, and also rapid strategies for straight drug testing. Contrasted to bulk approaches, by effectively controlling the geometries of the produced chip and the circulation rates of multiphase fluids, microfluidic modern technology makes it possible for the generation of very stable, consistent, mono dispersed particles with greater encapsulation performance. Given that the existing preclinical versions are inefficient drug displays for anticipating clinical outcomes, microfluidic systems might use a more rapid as well as affordable choice. The development of pharmaceutical logical methods represents one of the most significant aspects of medication growth. Recent advancements in micro fabrication and microfluidics can provide brand-new techniques for drug analysis, including medicine screening, active testing as well as the study of metabolic process. Microfluidic chip innovations, such as lab-on-a-chip innovation, three-dimensional (3D) cell society, organs-on-chip as well as bead methods, have actually all been created swiftly.

Multi-Marker Immunomagnetic Enrichment of Circulating Melanoma Cells.

Within the last decade, circulating tumor cells (CTCs) have emerged as a promising biomarker for prognostication, treatment monitoring, and detection of markers of treatment resistance, and their isolation can be used as a minimally invasive means of profiling tumors across multiple body sites. However, CTCs represent a minuscule fraction of the total circulating cells in a patient. Therefore, sensitive isolation methods are needed for the detection and downstream analysis of these cells. Herein we describe a sensitive method for melanoma CTC isolation using a multi-marker immunomagnetic bead method. This method has been purposely optimized to detect CTCs in melanoma patients.