Design, Synthesis, and Cellular Imaging Application of a Fluorescent Probe Based on Fluoride Ion-Induced Cyclization of Phenothiazine Derivatives.

Fluoride is both necessary and potentially harmful in excessive amounts, making its detection crucial. Fluorescent probes provide a sensitive and selective means for this purpose. In this study, we developed and synthesized a fluorescent probe for LDT using phenothiazine derivatives and aryl vinyl nitrile. Initially non-fluorescent, the probe undergoes a Si-O bond breakage in the presence of fluoride ions, resulting in the formation of a larger conjugated system and subsequent fluorescence emission. The probe exhibits superior selectivity and sensitivity towards fluoride ions, with a detection limit of 0.35 µM. Moreover, cellular imaging experiments demonstrated the probe's effectiveness in recognizing fluoride ions within HepG2 cells.

Development of carbazole-based fluorescent probe for highly sensitive application in fluoride ion detection.

Fluorine is a necessary element for human, which is closely related to life activities, such as metabolism of teeth and bone tissue. A small amount of fluoride ions can promote the strengthen of our body. However, a large amount of fluoride ions will damage the human immune system to produce organ diseases. Sensitive and rapid detection of fluoride ions has attracted great interests for researchers. In this work, a reactive fluorescent probe SCP for detection of fluoride ions with high quantum yield was designed and synthesized based on the carbazole ring. Subsequently, the photophysical properties of the probe SCP were carefully studied. At last, SCP performed 62.8% quantum yield in physiological condition, excellent ability of quantitative analysis, well selectivity, and distinguishing features for HepG2 cell imaging.

[Efficacy and Safety of Cladribine-based Intensified Conditioning Regimen in Hematopoietic Stem Cell Transplantation in Patients with High-Risk Acute Myeloid Leukemia].

OBJECTIVE: To investigate the efficacy, safety and the risk factors affecting prognosis of high-risk acute myeloid leukemia (AML) patients treated by cladribine-based intensified conditioning regimen. METHODS: The clinical data of 28 patients with high-risk AML treated by cladribine in combination with busulfan plus cyclophosphamide (BuCy) intensified conditioning regimen before allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Zhujiang Hospital, Southern Medical University from October 2016 to June 2020 were analyzed retrospectively. The overall survival (OS) rate, cumulative progression-free survival (PFS) rate, relapse rate, non-relapse mortality (NRM), regimen related toxicity (RRT) and risk factors affecting prognosis of the patients were analyzed. RESULTS: The 1-year OS and PFS of the patients after implantation was (78.8±8.6)% and (79.8±8.1)%, while the 1-year cumulative relapse rate and NRM of the patients was 9.3% and 22.0%, respectively. The 1-year expected OS of MRD- high-risk patients before HSCT was 100%. The 1-year expected OS and PFS of the patients in pre-transplant relapse group was (46.9±18.7)% and (50.0±17.7)%, respectively. The incidence of I/II grade RRT was 39.3%. NO III/IV grade RRT were found in 28 patients. Multivariate analysis showed that pre-transplant relapse was the independent risk factor affecting OS and PFS of the patients. CONCLUSION: The intensified conditioning regimen of cladribine in combination with BuCy can reduce the relapse rate of high-risk AML transplantation, and its RRT is mild, exhibiting good safety. MRD- high-risk patients before HSCT can achieve better transplant benefits, but the prognosis of patients with relapse before transplantation is not significantly improved. Therefore, for non-relapsed high-risk AML patients, this intensified conditioning regimen deserves to be considered.

A two-phase approach for the Radiotherapy Scheduling Problem.

The Radiotherapy Scheduling Problem (RTSP) focuses on optimizing the planning of radiotherapy treatment sessions for cancer patients. In this paper, we propose a two-phase approach for the RTSP. In the first phase, radiotherapy sessions are assigned to specific linear accelerators (linacs) and days. The second phase then decides the sequence of patients on each day/linac and the specific appointment times. For the first phase, an Integer Linear Programming (IP) model is proposed and solved using CPLEX. For the second phase, a Mixed Integer Linear Programming (MIP) and a Constraint Programming (CP) model are proposed. The test data is generated based on real data from CHUM, a large cancer center in Montréal, Canada, with an average of 3,500 new patients and 40,000 radiotherapy treatments per year. The results show that in the second phase, CP is better at finding good solutions quickly while MIP is better at closing optimality gaps with more run time. Lastly, a simulation is conducted to evaluate the impact of different scheduling strategies on the outcome of the scheduling. Preliminary results show that batch scheduling reduces patients' waiting time and overdue time.

The effects of NCBP3 on METTL3-mediated m6A RNA methylation to enhance translation process in hypoxic cardiomyocytes.

Hypoxia as a crucial pathogenesis factor usually results in huge harmful effects on cardiac injury and dysfunction. Our previous study has uncovered the global transcriptome and translatome profiles of cardiomyocytes in vitro and in vivo to response to hypoxia by RNA sequencing and ribosome profiling sequencing. We observe a series of differential expressed genes between transcription and translation, which may be attributed to the hypoxia-specific binding affinity of nuclear cap-binding subunit 3 (NCBP3) at 5' untranslation region of target genes. Although we observe that NCBP3 can facilitate translational process in myocardium under hypoxia stress, the underlying molecular mechanism of NCBP3 for gene translation modulation remains unclear. In this study, we performed NCBP3 immunoprecipitation for mass spectrum and found that METTL3 and eIF4A2 particularly interacted with NCBP3 in hypoxic rat H9C2 cardiomyocytes. Furthermore, we observed that METTL3-mediated N6-methyladenosine (m6A) methylation was elevated in hypoxia, but compromised by NCBP3 or METTL3 knockdown. Finally, we also demonstrated that NCBP3/METTL3/eIF4A2 regulatory axis plays a specific role in cardiomyocytes undergoing hypoxic stress. Taken together, we unmasked NCBP3, a novel hypoxia-specific response protein functions as a scaffold to coordinate METTL3 and eIF4A2 for enhancing gene translation by m6A RNA methylation in cardiomyocytes upon hypoxic stress.

Smartphone-Based Paper Microfluidic Immunoassay of Salmonella and E. coli.

Previous studies from our lab have created a simple procedure for single-cell count of bacteria on a paper chip platform using optical detection from a smartphone. The procedure and steps employed are outlined along with the lessons learned and details of certain steps and how the design was optimized. Smartphone optical detection is easy to use, low cost, and potentially field deployable, which can be useful for early and rapid detection of pathogens. Smartphone imaging of a paper microfluidic chip preloaded with antibody-conjugated particles provides an adaptable platform for detection of different bacterial targets. The paper microfluidic chip was fabricated with a multichannel design. Each channel was preloaded with either a negative control of bovine serum albumin (BSA) conjugated particles, anti-Salmonella Typhimurium-conjugated particles with varying amounts (to cover different ranges of assay), or anti-Escherichia coli-conjugated particles. Samples were introduced to the paper microfluidic chip using pipetting. Antigens of Salmonella Typhimurium traveled through the channel by capillary action confined within the paper fibers surrounded by the hydrophobic barrier. The paper channel was observed to act as a filter for unwanted particles and contaminants found in field samples. Serial dilutions of known concentrations of bacterial targets were also tested using this procedure to construct a standard curve prior to the assays. The antibody-conjugated particles were able to immunoagglutinate which was quantified through evaluation of Mie scatter intensity. This Mie scattering was quantified in images taken with a smartphone at an optimized angle and distance. Mie scatter simulation provided a method of optimizing the experimental setup and could translate easily to other types of target sample matrices. A smartphone application was developed to help the user position the smartphone optimally in relation to the paper microfluidic chip. The application integrated both image capturing capability and a simple image processing algorithm that calculated bacteria concentrations. The detection limit was at a single-cell level with a total assay time ranging from 90 to less than 60 s depending on the target.

[Efficacy and Prognosis of Allogeneic Hematopoietic Stem Cell Transplantation for Acute Monocytic Leukemia Patients].

OBJECTIVE: To investigate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of acute monocytic leukemia (AML-M5) and the related factors that affecting the prognosis of the patients. METHODS: The clinical data of 71 patients with AML-M5 treated with allo-HSCT in Zhujiang Hospital Affiliated to Southern Medical University from April 2009 to October 2019 were collected and retrospectively analyzed. The incidence of graft-versus-host disease (GVHD), cumulative overall survival (OS) rate, cumulative progression-free survival (PFS) rate, transplantation-related mortality (TRM), relapse rate and the risk factors affecting prognosis in the patients were analyzed. RESULTS: 66 patients obtained hematopoietic reconstruction after transplantation, the median time of granulocyte implantation was 12 (9-26) d, and the median time of megakaryocytic implantation was 13 (8-72) d. The incidence of acute GVHD and chronic GVHD was 33.8% (24/71) and 36.6% (26/71), respectively. The median follow-up time was 13.81 (0.16 to 112.54) months; the median OS and PFS was 31.27 and 26.07 months, respectively. The cumulative OS of the patients in 1 and 3 years after transplantation was 64.9% and 48.6%, respectively, and the cumulative PFS of the patients in 1 and 3 years was 55.0% and 39.5%, respectively. The cumulative relapse rate of the patients in 1 and 3 years was 24% and 40%, respectively. Multivariate analysis showed that pre-transplantation relapse was the independent risk factor affecting OS (HR=2.32, 95%CI:1.17-4.62, P=0.02) and PFS (HR=3.08, 95%CI:1.61-5.90, P=0.001) of the patients; invasive fungal disease after transplantation was the independent risk factor affecting OS (HR=2.71, 95% CI:1.32-5.56, P=0.007) and PFS (HR=2.87, 95%CI=1.40-5.86, P=0.004) of the patients; FLT3 mutation was the independent risk factor affecting PFS (HR=2.13, 95%CI=1.07-4.24, P=0.03) of the patients. CONCLUSION: AML-M5 is the intermediate or high-risk leukemia, and allo-HSCT can improve the survival prognosis of the patients. Pre-transplantation relapse and invasive fungal disease after transplantation are the important factors affecting the efficacy of allo-HSCT in patients with AML-M5.

Multi-Normalization and Interpolation Protocol to Improve Norovirus Immunoagglutination Assay from Paper Microfluidics with Smartphone Detection.

Norovirus (NoV) is one of the leading causes of acute gastroenteritis, affecting 685 million people per year around the world. The best preventive measure is to screen water for possible NoV contamination, not from infected humans, preferably using rapid and field-deployable diagnostic methods. While enzyme immunoassays (EIAs) can be used for such detection, the low infectious dose as well as the generally inferior sensitivity and low titer of available NoV antibodies render critical challenges in using EIAs toward NoV detection. In this work, we demonstrated smartphone-based Mie scatter detection of NoV with immunoagglutinated latex particles on paper microfluidic chips. Using only three different concentrations of anti-NoV-conjugated particles, we were able to construct a single standard curve that covered seven orders of magnitude of NoV antigen concentrations. Multiple normalization steps and interpolation procedures were developed to estimate the optimum amount of antibody-conjugated particles that matched to the target NoV concentration. A very low detection limit of 10 pg/mL was achieved without using any concentration or enrichment steps. This method can also be adapted for detection of any other virus pathogens whose antibodies possess low sensitivity and low antibody titer.

Smartphone Detection of UV LED-Enhanced Particle Immunoassay on Paper Microfluidics.

Use of a smartphone as an optical detector for paper microfluidic devices has recently gained substantial attention due to its simplicity, ease of use, and handheld capability. Utilization of a UV light source enhances the optical signal intensities, especially for the particle immunoagglutination assay that has typically used visible or ambient light. Such enhancement is essential for true assimilation of assays to field deployable and point-of-care applications by greatly reducing the effects by independent environmental factors. This work is the first demonstration of using a UV LED (UVA) to enhance the Mie scatter signals from the particle immunoagglutination assay on the paper microfluidic devices and subsequent smartphone detection. Smartphone's CMOS camera can recognize the UVA scatter from the paper microfluidic channels efficiently in its green channel. For an Escherichia coli assay, the normalized signal intensities increased up to 50% from the negative signal with UV LED, compared with the 4% to 7% with ambient light. Detection limit was 10 colony-forming units/mL. Similar results were obtained in the presence of 10% human whole blood.

[Clinical Summarization of Allogeneic Hematopoietic Stem Cell Transplantation for Leukemia: A Report of 100 Cases].

OBJECTIVE: To analyze the treatment outcome of a consecutive series of 100 leukemia patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: The clinical data of leukemia patients received allo-HSCT were analyzed retrospectively, the therapeutic efficacy was summarized. 100 evaluable cases of leukemia included 47 cases of AML, 33 cases of ALL, 2 cases of AL (biphenotypic), 16 CML and 2 CMML. Before transplantation, 76 cases were in first complete remission, 9 cases in second or greater complete remission and 15 cases in non-remission or relapse. All the patients received peripheral blood hematopoietic stem cell transplantation (PBHSCT). The conditioning regimen of human leukocyte antigen (HLA)-matched allo-HSCT group was modified BuCy, but in HLA-mismatched group Fludarabine and anti-human thymocyte globulin (ATG) was added. CsA+MTX regimen was used for prophylaxis of graft-versus-host disease (GVHD) in HLA-identical allo-HSCT, while additional MMF was added in HLA-mismatched group. The average time of follow-up was 13 months. RESULTS: At the last follow-up, 66.0% (66/100) patients survived, 53.0% (53/100) patients survived without leukemia, 28.0% (28/100) patients relapsed and 34.0% (34/100) patients died, 44.1% patients of them died from infectious pulmonary complications. During transplantation, 65.0% of the patients were suffered from lung infection. The overall survival (OS) and disease-free survival (DFS) of all cases was 60.9% and 48.8%, respectively. The recurrence rate was significantly higher in non-remission (66.7%) than in CR (21.2%) patients (P < 0.05). The cumulative incidence of GVHD in HLA-mismatched transplantation was 60.8%, which was significantly higher than that of HLA-matched transplantation (38.8%) (P < 0.05). CONCLUSION: Allo-HSCT can cure a significant proportion of leukemia patients, especially for those in CR status. Since the incidence of infectious pulmonary complications after allo-HSCT is still high, much more attention should be paid to it. The comprehensive prognosis of HLA-matched transplantation is better than the HLA-mis-matched transplantation.

[Research Progres on Relationship between the Graft-Versus-Host Disease and Cytomegalovirus Infection--Review].

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of leukemia patients can improve overall survival and disease-free survival, and reduce relapse. Although the allo-HSCT is more widely used in the treatment of leukemia, but the graft-versus-host disease (GVHD) and cytomegalovirus (CMV) infections are the common complications, and are the major cause of mortality for patients following allo-HSCT. Previous studies showed that there might be a mutual promotive relationship between GVHD and CMV infection, but the clear relationship remained to be elucidated. The relationship of GVHD and CMV has been the focus of clinical research. Recently, a great progress has been made on researches of the relationship and its mechanism between GVHD and CMV infection. In this article, the relationship and its mechanism between GVHD and CMV infection after allo-HSCT are reviewed.

[Progress of Clinical Trials on Bone Marrow Mesenchymal Stem Cells for Prevention and Therapy of Graft-Versus-Host Disease].

Graft-versus-host disease (GVHD) is a major complication following allogenetic hematopoietic stem cell transplantation, which shows a great threat to patients' survival and life quality. Along with multiple differentiation potential to various types of progenitor cells, bone marrow mesenchymal stem cells (BMMSC) have been confirmed to possess low immunogenicity and exert favorable immunomodulation. The recent studies show that the safety and high efficiency of BMMSC to prevent and cure GVHD greatly improved survival rate of the hosts. The most recent progress on prevention and therapy of GVHD is summarized in this review based on biology of BMMSC and pathogenesis of GVHD, so as to provide the effective evidence for further research.

Colorimetric Sensor Array for White Wine Tasting.

A colorimetric sensor array was developed to characterize and quantify the taste of white wines. A charge-coupled device (CCD) camera captured images of the sensor array from 23 different white wine samples, and the change in the R, G, B color components from the control were analyzed by principal component analysis. Additionally, high performance liquid chromatography (HPLC) was used to analyze the chemical components of each wine sample responsible for its taste. A two-dimensional score plot was created with 23 data points. It revealed clusters created from the same type of grape, and trends of sweetness, sourness, and astringency were mapped. An artificial neural network model was developed to predict the degree of sweetness, sourness, and astringency of the white wines. The coefficients of determination (R2) for the HPLC results and the sweetness, sourness, and astringency were 0.96, 0.95, and 0.83, respectively. This research could provide a simple and low-cost but sensitive taste prediction system, and, by helping consumer selection, will be able to have a positive effect on the wine industry.

Smartphone-based, sensitive µPAD detection of urinary tract infection and gonorrhea.

The presence of bacteria in urine can be used to monitor the onset or prognosis of urinary tract infection (UTI) and some sexually-transmitted diseases (STDs), such as gonorrhea. Typically, bacteria's presence in urine is confirmed by culturing samples overnight on agar plates, followed by a microscopic examination. Additionally, the presence of Escherichia coli in a urine sample can be indirectly confirmed through assaying for nitrite (generated by reducing nitrate in urine), however this is not sufficiently specific and sensitive. Species/strains identification of bacteria in a urine sample provides insight to appropriate antibiotic treatment options. In this work, a microfluidic paper analytical device (µPAD) was designed and fabricated for evaluating UTI (E. coli) and STD (Neisseria gonorrhoeae) from human urine samples. Anti-E. coli or anti-N. gonorrhoeae antibodies were conjugated to submicron particles then pre-loaded and dried in the center of each paper microfluidic channel. Human urine samples (undiluted) spiked with E. coli or N. gonorrhoeae were incubated for 5 min with 1% Tween 80. The bacteria-spiked urine samples were then introduced to the inlet of paper microfluidic channel, which flowed through the channel by capillary force. Data confirms that proteins were not filtered by µPAD, which is essential for this assay. Urobilin, the component responsible for the yellow appearance of urine and green fluorescence emission, was filtered by µPAD, resulting in significantly minimized false-positive signals. This filtration was simultaneously made during the µPAD assay and no pretreatment/purification step was necessary. Antibody-conjugated particles were immunoagglutinated at the center of the paper channel. The extent of immunoagglutination was quantified by angle-specific Mie scatter under ambient lighting conditions, utilizing a smartphone camera as a detector. The total µPAD assay time was less than 30s. The detection limit was 10 CFU/mL for both E. coli and N. gonorrhoeae, while commercially available gonorrhea rapid kit showed a detection limit of 10(6) CFU/mL. A commercially available nitrite assay test strip also had a detection limit of 10(6) CFU/mL, but this method is not antibody-based and thus not sufficiently specific. By optimizing the particle concentration, we were also able to extend the linear range of the assay up to 10(7) CFU/mL. The proposed prototype will serve as a low-cost, point-of-care, sensitive urinalysis biosensor to monitor UTI and gonorrhea from human urine.

Overexpression of EPS8 is associated with poor prognosis in patients with acute lymphoblastic leukemia.

Molecular markers have become an invaluable tool in monitoring disease status particularly of leukemias, as bone marrow samples can be easily collected for analysis during all stages of disease development including diagnosis, treatment, and follow-up. Two genes that have been used as prognostic markers in acute leukemia are Wilms' tumor (WT1) and multidrug resistance-1 (MDR1). A novel gene, epidermal growth factor receptor pathway substrate 8 (EPS8), is often over-expressed and associated with poor outcome in some solid tumor types. However, whether EPS8 is also associated with the development of acute lymphoblastic leukemia (ALL) is unclear. Here, quantitative real-time PCR was used to evaluate the expression of EPS8, MDR1, and WT1 in bone marrow samples of adult ALL patients (n=107) and non-leukemia controls (n=22). EPS8, MDR1, and WT1 were detected in ALL patients, and significant correlations were found between expression profiles for EPS8 and MDR1, EPS8 and WT1, and MDR1 and WT1. In general, high expression of EPS8, MDR1, or WT1 in patients was associated with a higher risk of relapse. Furthermore, when patients were stratified based on high or low expression of the genes, Kaplan-Meier survival analysis indicated that disease-free survival of patients with the high-EPS8/high-WT1/high-MDR1 profile was significantly shorter than in patients with the low-EPS8/low-WT1/low-MDR1 profile or those excluded from either of these groups (P<0.0001). Thus, EPS8, as MDR1 and WT1, may be a clinically valuable biomarker for assessing the outcome of ALL patients.

Rapid and reagentless detection of microbial contamination within meat utilizing a smartphone-based biosensor.

A smartphone-utilized biosensor was developed for detecting microbial spoilage on ground beef, without using antibodies, microbeads or any other reagents, towards a preliminary screening tool for microbial contamination on meat products, and potentially towards wound infection. Escherichia coli K12 solutions (10(1)-10(8) CFU/mL) were added to ground beef products to simulate microbial spoilage. An 880 nm near infrared LED was irradiated perpendicular to the surface of ground beef, and the scatter signals at various angles were evaluated utilizing the gyro sensor and the digital camera of a smartphone. The angle that maximized the Mie scatter varied by the E. coli concentration: 15° for 10(8) CFU/mL, 30° for 10(4) CFU/mL, and 45° for 10 CFU/mL, etc. SEM and fluorescence microscopy experiments revealed that the antigens and cell fragments from E. coli bonded preferably to the fat particles within meat, and the size and morphologies of such aggregates varied by the E. coli concentration.

Smartphone-based optofluidic lab-on-a-chip for detecting pathogens from blood.

A novel smartphone-based detection device was created to detect infectious pathogens directly from diluted (10%) human whole blood. The model pathogen was histidine-rich protein 2 (HRP-2), an antigen specific to Plasmodium falciparum (malaria). Anti-HRP-2-conjugated submicrobeads were mixed with HRP-2-infused 10% blood in a lab-on-a-chip device. The white LED flash and the digital camera of the smartphone were used as light source and detector, which delivered light to and from the bead and blood mixture via optofluidic channels in the lab-on-a-chip. The optofluidic channels were angled at 45 degrees to capture the Mie scatter from the sample. Considering the absorption and scattering characteristics of blood (red/infrared preferred) and the Mie scatter simulations for microbead immunoagglutination (UV preferred), blue detection showed the best results. The detection limit was 1 pg/mL in 10% blood. The linear range was from 1 pg/mL to 10 ng/mL. A handheld device, easily attachable to a single smartphone, was finally designed and fabricated using optical mirrors and lenses and successfully detected the HRP-2 from 10% blood. The total assay time was approximately 10 min. The proposed device can potentially be used for detecting a wide range of blood infection with high sensitivity.

Enhanced nucleic acid amplification with blood in situ by wire-guided droplet manipulation (WDM).

There are many challenges facing the use of molecular biology to provide pertinent information in a timely, cost effective manner. Wire-guided droplet manipulation (WDM) is an emerging format for conducting molecular biology with unique characteristics to address these challenges. To demonstrate the use of WDM, an apparatus was designed and assembled to automate polymerase chain reaction (PCR) on a reprogrammable platform. WDM minimizes thermal resistance by convective heat transfer to a constantly moving droplet in direct contact with heated silicone oil. PCR amplification of the GAPDH gene was demonstrated at a speed of 8.67 s/cycle. Conventional PCR was shown to be inhibited by the presence of blood. WDM PCR utilizes molecular partitioning of nucleic acids and other PCR reagents from blood components, within the water-in-oil droplet, to increase PCR reaction efficiency with blood in situ. The ability to amplify nucleic acids in the presence of blood simplifies pre-treatment protocols towards true point-of-care diagnostic use. The 16s rRNA hypervariable regions V3 and V6 were amplified from Klebsiella pneumoniae genomic DNA with blood in situ. The detection limit of WDM PCR was 1 ng/µL or 10(5)genomes/µL with blood in situ. The application of WDM for rapid, automated detection of bacterial DNA from whole blood may have an enormous impact on the clinical diagnosis of infections in bloodstream or chronic wound/ulcer, and patient safety and morbidity.

Smartphone quantifies Salmonella from paper microfluidics.

Smartphone-based optical detection is a potentially easy-to-use, handheld, true point-of-care diagnostic tool for the early and rapid detection of pathogens. Paper microfluidics is a low-cost, field-deployable, and easy-to-use alternative to conventional microfluidic devices. Most paper-based microfluidic assays typically utilize dyes or enzyme-substrate binding, while bacterial detection on paper microfluidics is rare. We demonstrate a novel application of smartphone-based detection of Salmonella on paper microfluidics. Each paper microfluidic channel was pre-loaded with anti-Salmonella Typhimurium and anti-Escherichia coli conjugated submicroparticles. Dipping the paper microfluidic device into the Salmonella solutions led to the antibody-conjugated particles that were still confined within the paper fibers to immunoagglutinate. The extent of immunoagglutination was quantified by evaluating Mie scattering from the digital images taken at an optimized angle and distance with a smartphone. A smartphone application was designed and programmed to allow the user to position the smartphone at an optimized angle and distance from the paper microfluidic device, and a simple image processing algorithm was implemented to calculate and display the bacterial concentration on the smartphone. The detection limit was single-cell-level and the total assay time was less than one minute.

[Cytotoxicity of Naja Naja Actra Venom Component combined with activated immune cells on leukemia cell line KG1a].

This study was aimed to investigate the cytotoxic effect of the Naja Naja Actra Venom Component (NNAVC) combined with activated immune cells on human acute myeloblastic leukemia line KG1a cells. The cytotoxic effects of NNAVC at different concentrations on KG1a cells were measured by CCK-8 method. LDH releasing assay was used to detect the cytotoxic effects of activated immune cells, NNAVC combined with activated immune cells on KG1a cells and the sensitivity of KG1a treated with NNAVC to activated immune cells. The results showed that the inhibitory rate of NNAVC on KG1a cells increased with the concentration enhancement, the cytotoxicity of activated immune cells at the different effector to target (E:T) ratios(6.25:1, 12.5:1, 25:1) on KG1a cells were 12.30%, 24.85% and 52.26%. The cytotoxicity of NNAVC combined with activated immune cells at the different E:T cell ratios (10:1, 20: 1) on KG1a cells were 56.21% and 85.59%, which were higher than that of NNAVC or activated immune cells alone. The cytotoxicity of activated immune cells at the E: T cell ratio of 10:1 on KG1a cells treated with NNAVC at different concentrations were 25.65%, 31.33%, 28.63% and 16.78%, respectively, and that at the E:T cell ratio of 20: 1 were 40.62%, 44.70%, 44.62% and 40.72%. It is concluded that:both of NNAVC and activated immune cells have lethal effect on KG1a cells, and the combination of NNAVC and activated immune cells can strengthen their effect on KG1a.