Brucellosis is a bacterial infectious disease caused mainly by Brucella. Transmission is mainly by contact with infected domestic or wild animals or their excreta. Clinical diagnosis of brucellosis is usually based on qualitative total antibody tests, which make it difficult to differentiate between acute and pre-existing infections. In this study, for the simultaneous detection of anti-brucellosis IgG and IgM, we innovatively developed a surface-enhanced Raman scattering (SERS)-lateral flow immunoassay (LFIA) detection system, and evaluated its performance and diagnostic effect by using clinical serum samples. The key design of this biosensor involves the preparation of two immunoprobes using two Raman spheres (R-Sphere) with distinct signals conjugated to mouse anti-human IgG and IgM, respectively. In this design, brucellosis-specific antigens are embedded in the T-line, allowing simultaneous capture of anti-brucellosis IgG and IgM in a specific binding reaction. A portable Raman instrument is used to detect the characteristic signal intensities generated by IgM and IgG on the T-line, which can then be used to analyze the target IgM and IgG antibodies. Based on the analysis of 40 clinical samples, the method has a sensitivity and specificity of 100%, and the detection time is only 15 min. The advantages of this technology are fast speed, convenient use, high sensitivity, and it can distinguish the disease course to achieve precise treatment. These features make it an ideal solution for large-scale brucellosis testing in remote and nomadic areas, where it can play a crucial role in improving public health protection. Brucellosis is a bacterial infectious disease caused mainly by Brucella. Transmission is mainly by contact with infected domestic or wild animals or their excreta.

The present study sought to develop a cardiac troponin I (cTnI) detection system based on background fluorescence quenching of internal filtration effect (IFE) and study the influence of IFE on the sensitivity of cTnI detection. Three nanogold materials were synthesized as fluorescence quenchers, and rhodamine 6 G (R6G) and Cy5 were used as fluorescence probes. Six experimental systems were established to detect cTnI in negative serum test solutions and clinical serum samples. The sensitivity of each system was compared to explore the contribution of IFE to the detection sensitivity of cTnI. When applied to negative serum test solutions, the R6G-nanogold material I system exhibited a superior detection effect for cTnI, with a limit of detection (LOD) of 0.15 ng ml-1. When applied to clinical serum samples, the Cy5-nanogold material Ⅲ system yielded a better detection effect for cTnI, with the lowest concentration of cTnI detected at 2 ng ml-1. The first and second internal filtering effects in the proposed system can be achieved simultaneously, effectively avoiding light absorption interference from clinical serum samples and enhancing the sensitivity of the background fluorescence quenching detection of cTnI.

Background: Since the poor response to existing anti-tuberculosis drugs and low drug concentration in local bone tissues, the traditional drug therapy does not result in satisfactory treatment of osteoarticular tuberculosis. Thus, we report a rifapentine release system with imparted bone targeting potential using tetracycline (TC) -modified nanoparticles (NPs).Methods: TC was conjugated to PLGA-PEG copolymer via a DCC/NHS technique. Rifapentine-loaded NPs were prepared by premix membrane emulsification technique. The resulting NPs were characterized in terms of physicochemical characterization, hemolytic study, cytotoxicity, bone mineral binding ability, in vitro drug release, stability test and antitubercular activity. The pharmacokinetic and biodistribution studies were also performed in mice.Results: Rifapentine loaded TC-PLGA-PEG NPs were proved to be 48.8 nm in size with encapsulation efficiency and drug loading of 83.3% ± 5.5% and 8.1% ± 0.4%, respectively. The release of rifapentine from NPs could be maintained for more than 60 h. Most (68.0%) TC-PLGA-PEG NPs could bind to HAp powder in vitro. The cellular studies revealed that NPs were safe for intravenous administration. In vivo evaluations also revealed that the drug concentration of bone tissue in TC–PLGA–PEG group was significantly higher than that in other groups at all time (p < 0.05). Both NPs could improve pharmacokinetic parameters without evident organ toxicity. The minimal inhibitory concentration of NPs was 0.094 μg/mL, whereas this of free rifapentine was 0.25 μg/mL.Conclusion: Rifapentine loaded TC-PLGA-PEG NPs could increase the amount of rifapentine in bone tissue, prolong drug release in systemic circulation, enhance anti-tuberculosis activity, and thereby reducing dose and frequency of drug therapy for osteoarticular tuberculosis.

Inflammation is a key factor in the development of ulcerative colitis (UC). 1,25-dihydroxyvitamin D3 (1,25(OH)2D3, VD3), as the major active ingredient of vitamin D and an anti-inflammatory activator, is closely related to the initiation and development of UC, but its regulatory mechanism remains unclear. In this study, we carried out histological and physiological analyses in UC patients and UC mice. RNA sequencing (RNA-seq), assays for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq), chromatin immunoprecipitation (ChIP) assays and protein and mRNA expression were performed to analyze and identify the potential molecular mechanism in UC mice and lipopolysaccharide (LPS)-induced mouse intestinal epithelial cells (MIECs). Moreover, we established nucleotide-binding oligomerization domain (NOD)-like receptor protein nlrp6-/- mice and siRNA-NLRP6 MIECs to further characterize the role of NLRP6 in anti-inflammation of VD3. Our study revealed that VD3 abolished NOD-like receptor protein 6 (NLRP6) inflammasome activation, suppressing NLRP6, apoptosis-associated speck-like protein (ASC) and Caspase-1 levels via the vitamin D receptor (VDR). ChIP and ATAC-seq showed that VDR transcriptionally repressed NLRP6 by binding to vitamin D response elements (VDREs) in the promoter of NLRP6, impairing UC development. Importantly, VD3 had both preventive and therapeutic effects on the UC mouse model via inhibition of NLRP6 inflammasome activation. Our results demonstrated that VD3 substantially represses inflammation and the development of UC in vivo. These findings reveal a new mechanism by which VD3 affects inflammation in UC by regulating the expression of NLRP6 and show the potential clinical use of VD3 in autoimmune syndromes or other NLRP6 inflammasome-driven inflammatory diseases.

In this study, by comparing the UV-vis spectral characteristics of colloidal gold and colloidal gold enhancer, and their differences as immunochromatographic tracers in the qualitative detection of PCT, IL-6, Hp and quantitative determination of PCT performance, the factors that may affect the sensitivity were discussed. The results show that the absorbance at 520 nm of CGE diluted 20-fold and colloidal gold diluted 2-fold were comparable, and the sensitivity of CGE immunoprobe for qualitative detection of PCT, IL-6 and Hp was higher than that of colloidal gold immunoprobe, and the reproducibility and accuracy of both immunoprobes for quantitative detection of PCT were good. Indicating that the high sensitivity of CGE immunoprobe detection is mainly due to the absorption coefficient of CGE at 520 nm is about 10 times that of colloidal gold immunoprobe, CGE has stronger light absorption capacity and stronger quenching effect on rhodamine 6 G on the nitrocellulose membrane surface of the test strip.

BackgroundMyeloid Sarcoma with monocytic differentiation is rare and quite likely is missed by surgical pathologists. However it is frequently misdiagnosed because of its non-specific imaging and histological pattern.Case presentationWe report the case of a 64-year-old woman with gastric primary myeloid sarcoma with monocytic differentiatio. Upper endoscopy revealed a neoplastic growth at the junction of the lesser curvature and gastric antrum. Except for a slightly increased peripheral monocyte count, no abnormalities were found on hematological and bone-marrow examination. Gastroscopic biopsy showed poorly differentiated atypical large cells with visible nucleoli and nuclear fission. Immunohistochemistry showed positive CD34, CD4, CD43, and CD56 expression, and weakly positive lysozyme expression. Immune markers for poorly differentiated adenocarcinoma, malignant melanoma, and lymphohematopoietic-system tumors were negative. The final diagnosis was myeloid sarcoma with monocytic differentiation. Chemotherapy did not shrink the tumor, so, radical surgery was performed. Although the tumor morphology did not change postoperatively, the immunophenotype did. CD68 and lysozyme expression (tumor tissue markers) changed from negative and weakly positive to strongly positive, AE1/3 expression (epithelial marker) changed from negative to positive, and CD34, CD4, CD43, and CD56 expression (common in naive hematopoietic cell-derived tumors) was greatly attenuated. Exome sequencing revealed missense mutations in FLT3 and PTPRB, which are associated with myeloid sarcoma, and in TP53, CD44, CD19, LTK, NOTCH2, and CNTN2, which are associated with lymphohematopoietic tumors and poorly differentiated cancers.ConclusionWe diagnosed myeloid sarcoma with monocytic differentiation after excluding poorly differentiated adenocarcinoma, common lymphohematopoietic-system tumors, epithelioid sarcoma, and malignant melanoma. We identified that the immunophenotypic of patient had alterations after chemotherapy, and FLT3 gene mutations. We hope that the above results will improve our understanding of this rare tumor.

Background:Adriamycin (doxorubicin) is an important traditional drug that exhibits cytotoxicity in Diffuse Large B-cell Lymphoma (DLBCL). Doxorubicin affects the DLBCL cells at all stages of their cell cycle. Combined with our previous results, this study discovered that the overexpression of hsa-miR-28-5p inhibited the proliferation, promoted apoptosis, and triggered cell cycle arrest at the S-phase in DLBCL cells. However, the effect of (Homo sapiens, hsa)-microRNA (miR)-28-5p on doxorubicin sensitivity in DLBCL has not been investigated. This study aims to reveal the effects of hsa-miR-28-5p on doxorubicin sensitivity at the level of DLBCL cells.Methods:To determine the optimal concentration of doxorubicin, different concentrations of doxorubicin were used to treat DLBCL cells. CCK-8 assay was used to detect the proliferation of DLBCL cells. The hsa-miR-28-5p-mimic NC and hsa-miR-28-5p mimic were transfected to doxorubicin-mediated DLBCL cells. Simultaneously, blank control groups were set up. The cells were cultured and transfected for 24 h. Next, each group was administered with different concentrations of doxorubicin and cultured again for 24 h to observe the effects of hsa-miR-28-5p on doxorubicin sensitivity at different times. The proliferation, early apoptosis, and late apoptosis in DLBCL cells were determined using soft agar colony-forming assay, mitochondrial membrane potential assay, and caspase-3 activity assay, respectively. The apoptosis and cell cycle were explored using Annexin V-PE/7-AAD and PI/RNase staining buffer, respectively. We speculated that PD-L1 might be involved in the effect of hsa-miR-28-5p on the sensitivity of adriamycin (doxorubicin) in the DLBCL cells. Hence, we performed immunohistochemistry (IHC) to determine PD-L1 expression within formalin-fixed paraffin-embedded (FFPE) samples from 52 DLBCL cases.Results:The optimal concentration of doxorubicin targeting DLBCL cells was found to be 3.028 μmol/l. The effect of doxorubicin on DLBCL cells was time- and concentration-dependent. hsa-miR-28-5p mimic + doxorubicin remarkably decreased proliferation of DLBCL. DLBCL cell apoptosis rate was the highest in hsa-miR-28-5p mimic + doxorubicin group. Apart from that, hsa-miR-28-5p mimic plus doxorubicin had the best effect in promoting DLBCL cell apoptosis. After the intervention of hsa-miR-28-5p mimic + doxorubicin on DLBCL cells, the cell cycle was arrested in the S-phase and DNA synthesis was blocked. hsa-miR-28-5p mimic + doxorubicin could regulate the cycle of DLBCL cells. As a result, overexpression of hsa-miR-28-5p combined with doxorubicin is possibly involved in the development of DLBCL by affecting the proliferation, apoptosis, and cycle of DLBCL cells. PD-L1 showed an association with the prognosis of DLBCL patients. Combining with the literature, this suggested hsa-miR-28-5p may influence DLBCL occurrence and therapeutic effect by regulating the PD-L1 level.Conclusion:The combination of hsa-miR-28-5p mimic and doxorubicin may be considered more effective in inhibiting growth, arresting the cell cycle, and promoting cell apoptosis of DLBCL cells compared to using doxorubicin alone. The effects of doxorubicin on DLBCL cells were found to be time- and concentration-dependent. The overexpression of hsa-miR-28-5p enhanced the effect of doxorubicin on DLBCL cells, which may be attributed to the regulation of PD-L1 levels.

This study aimed to investigate the expression and prognostic significance of the signal transducer and activator of transcription protein 6 (STAT6YE361) and EB virus encoding a small molecule RNA (EBER) in Hodgkin lymphoma (HL), as well as their correlation with clinical parameters. The expression of STAT6YE361 and EBER was investigated in HL via immunohistochemistry and in situ hybridization. Patient clinical data were retrospectively collected from archival libraries, and statistical analysis was performed. Overall, the nuclear positive expression rate of STAT6YE361 was 46%, and the EBER positive expression rate was 57%. STAT6YE361 was specifically expressed on the nucleus in cHL tissues. EBER was overexpressed in HL and had correlations with several clinical data, including age, gender, ethnicity, and primary cancer site. Interestingly, nuclear STAT6YE361 expression was correlated with EBER expression. Based on survival analysis, the nuclear expression of STAT6YE361 and female patients were associated with poor prognosis and were independent prognostic factors for five-year OS. These findings suggest that STAT6YE361 is a potential valuable index in the differential diagnosis and prognosis of HL. The mechanism of STAT6YE361 is related to Epstein-Barr virus infection.

Aims A growing research demonstrated that YAP1 played important roles in gliomagenesis. We explored the expression of YAP1 and STAT3, the relationship between them and the effect of YAP1, STAT3 on prognosis in glioma. Methods Expression of YAP1, p-YAP1, STAT3, pSTAT3-S727 and pSTAT3-Y705 in 141 cases of low-grade gliomas (LGG) and 74 cases of high-grade gliomas (HGG) of surgical specimens were measured by immunohistochemistry. Pearson’s X2 test was used to determine the correlation between immunohistochemical expressions and clinicopathological parameters. Pearson’s or Spearman correlation test was used to determine the association between these proteins expression. Survival analysis was used to investigate the effect of these proteins on prognosis. Results High expressions of YAP1, STAT3, pSTAT3-S727 and pSTAT3-Y705 were found in HGG compared with LGG (p=0.000). High expressions of YAP1, STAT3, pSTAT3-S727 and pSTAT3-Y705 were found in 63.5%, 59.5%, 66.2% and 31.1% cases of HGG, respectively. YAP1 expression was associated to tumour location, Ki-67 and P53, STAT3 expression was related with Ki-67 and P53, and the expression of pSTAT3-S727 was associated with Ki-67. There was a significantly positive correlation between YAP1 and pSTAT3-S727 (p<0.0001; r=0.5663). Survival analysis revealed that patients with YAP1 and pSTAT3-S727 coexpression had worse overall survival (OS) and progression-free survival (PFS) (p<0.0001). Tumour grade, age, Ki-67 and YAP1 expression were independent prognostic factors for OS. In LGG group, both YAP1 and pSTAT3-S727 expressions were negative correlation with IDH1 mutation, YAP1 and pSTAT3-S727 coexpression showed worse OS and PFS of glioma patients. Conclusion Our research showed that YAP1 and STAT3 were significantly activated in HGG compared with LGG. YAP1 significantly correlated with pSTAT3-S727 in glioma, YAP1 and pSTAT3-S727 coexpression may serve as a reliable prognostic biomarker and therapeutic target for glioma.

The purpose of this investigation was to establish a mathematical model for determining the dissolution of diclofenac sodium and codeine phosphate simultaneously. Based on the dual-wavelength isosbestic point spectrophotometry, the dissolution of diclofenac sodium and codeine phosphate tablets was determined using Fiber-Optic Dissolution Test (FODT) instrument capable of real-time measurement. Dissolution curves showed that the dissolution process of diclofenac sodium was similar to that of codeine phosphate. The dissolution profile of diclofenac sodium and codeine phosphate at 45 min was concordant with that stated in Chinese pharmacopoeia. There was no significant difference between results obtained from FODT and HPLC (p>0.05). A fibre-optic dissolution test system assisted by the mathematical separation model of linear equations was able to detect the dissolution of diclofenac sodium and codeine phosphate simultaneously. The dissolution profiles and overall data, which can directly reflect the dissolution speed at each time point, can provide the basis for establishing standards for the quality evaluation of drugs.

Background The tumor suppressor protein p53 is a most promising target for the development of anticancer drugs. Allicin (diallylthiosulfinate) is one of the most active components of garlic ( Alliium sativum L. ) and possesses a variety of health-promoting properties with pharmacological applications. However, whether allicin plays an anti-cancer role against breast cancer cells through the induction of p53-mediated apoptosis remains unknown. Methods and results In this study, we investigate the anti-breast cancer effect of allicin in vitro by using MCF-7 and MD-MBA-231 cells. We found that allicin reduces cell viability, induces apoptosis and cell cycle arrest in both cells. Allicin activated p53 and caspase 3 expressions in both cells but produced different effects on the expression of p53-related biomarkers. In MDA-MB-231 cells, allicin up-regulated the mRNA and protein expression of A1BG and THBS1 while down-regulated the expression of TPM4. Conversely, the mRNA and protein expression of A1BG, THBS1 and TPM4 were all reduced in MCF-7 cells. Hence, allicin induces cell cycle arrest and apoptosis in breast cancer cells through p53 activation but it effects on the expression of p53-related biomarkers were dependent upon the specific type of breast cancer involved. Conclusions These findings suggest that allicin induces apoptosis and regulates biomarker expression in breast cancer cell lines through modulating the p53 signaling pathway. Furthermore, our results promote the utility of allicin as compound for further studies as an anticancer drug targeting p53.

We aimed to screen and identify differentially expressed microRNAs (miRNAs) between diffuse large B-cell lymphoma (DLBCL) and control (lymph node reactive hyperplasia, LRH) groups, investigate whether miRNAs associated with DLBCL and could serve as potential therapeutic targets. Five DLBCL experimental samples and five control samples were obtained from fresh tissues. The exclusion criteria were expressed as follows: patients with other lymphoid diseases and patients undergoing chemical treatment. Firstly, fresh samples were analyzed using microRNA microarray to identify differentially expressed miRNAs. Next, three databases (Targetscan, microRNAorg, PITA) were used to intersection predict the potential target gene of the 204 differential miRNAs and performed Venn diagram of the results. Then, target genes of differential miRNAs were performed by Gene ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway analysis. Finally, to validate the microRNA microarray data, we performed Real-time reverse transcription PCR (qRT-PCR) for eight differentially expressed miRNAs (miR-193a-3p, miR-19a-3p, miR-19b-3p, miR-370-3p, miR-1275, miR-490-5p, miR-630 and miR-665) using DLBCL and LRH fresh samples. Two hundred and four miRNAs showed differential expression, including 105 down-regulated and 54 up-regulated miRNAs. The cut-off criterion was set as P value ≤0.05 and fold change (FC) ≥2. A total of 7522 potential target genes for the 204 miRNAs were predicted. Potential target genes were enriched in pathway in cancers, MAPK signal pathway, regulation of actin cytoskeleton, focal adhesion, endocytosis, Wnt signal pathway, axon guidance, calcium signal pathway, and PI3K/AKT signal pathway. Eight miRNAs validation by qRT-PCR, four miRNAs (miR-19b-3p, miR-193a-3p, miR-370-3p and miR-490-5p) were low expression in DLBCL (P< 0.05). The high expression of miR-630 in DLBCL was statistically significant (P< 0.05). Our results of identification of differentially expressed miRNAs, predicted the target gene of the differential miRNAs, enrichment of target gene biological functions and signaling pathway, will provide basis for researchers in identifying the pathogenesis of DLBCL, could serve as reliable biomarkers for precise diagnosis and as therapeutic target for improvement of therapeutic efficacy in DLBCL in the future.

BACKGROUND:Tuberculosis (TB) is a leading cause of death amongst infectious diseases. The poor response to antitubercular agents necessitates the long-term use of high drug doses, resulting in low patient compliance, which is the main reason for chemotherapy failure and contributes to the development of multidrug-resistant TB. Patient non-compliance has been a major obstacle in the successful management of TB. The aim of this work was to develop and characterise rifapentine (RPT)-loaded PLGA-based nanoparticles (NPs) for reducing dosing frequency.METHODS:RPT-loaded PLGA and PLGA-PEG NPs were prepared using premix membrane homogenisation combined with solvent evaporation method. The resulting NPs were characterised in terms of physicochemical characteristics, toxicity, cellular uptake and antitubercular activity. NPs were further evaluated for pharmacokinetic and biodistribution studies in mice.RESULTS:The resulting NPs showed suitable and safe physicochemical characteristics and could be taken up by macrophages. RPT-loaded NPs were more effective against Mycobacterium tuberculosis than free RPT. In vivo studies revealed that NPs could improve pharmacokinetic parameters, particularly for RPT/PLGA-PEG NPs. Moreover, both formulations had no toxicity to the organs of mice and could reduce hepatotoxicity.CONCLUSION:The application of PLGA-based NPs as sustained-release delivery vehicles for RPT could prolong drug release, modify pharmacokinetics, increase antitubercular activity and diminish toxicity, thereby allowing low dosage and frequency.

This study aimed to investigate the hub protein related to the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway in diffuse large B-cell lymphoma (DLBCL). We used proteomics methods (iTRAQ) to explore the differentially expressed proteins in the non-germinal center B-cell -like (non-GCB) DLBCL in our previous study. In this study, a total of 137 formalin-fixed paraffin-embedded DLBCL tissue samples were analyzed via immunohistochemistry to verify the expression of TCL1, AKT1 + 2+3, IKKβ and to determine the differentially expressed proteins associated with the PI3K/AKT signaling pathway. Spearman correlation was used to analyze the relationship between these proteins, and survival analysis was used to investigate their effects on prognosis. Immunohistochemistry analysis indicated that TCL1, AKT1 + 2+3, and IKKβ were highly positively expressed in DLBCL. Results showed that the expression of TCL1 was related to ethnicity (p = 0.022), primary site (p = 0.045), Ann Arbor stage (p = 0.037), the International Prognostic Index (p = 0.005), β2-microglobulin (p = 0.030), BCL2 expression (p < 0.001), and Ki-67 expression (p = 0.008). A positive correlation was found between TCL1 and AKT1 + 2+3 (p < 0.001; r = 0.475). A positive correlation was also found between AKT1 + 2+3 and IKKβ (p < 0.001; r = 0.342). In survival analysis, anemia, non-treatment with R‑CHOP, positive TCL1 expression, and Ki-67 expression≥50% independently predicted short progression-free survival and overall survival in the total cohort (p < 0.05). Thus, TCL1 as a hub protein is associated with the PI3K/AKT signaling pathway in DLBCL. TCL1 expression indicated a poor prognosis in patients with DLBCL. With further studies, TCL1 may be established as a reliable prognostic biomarker and potential immunotherapeutic target for improving therapeutic efficacy for DLBCL in the future.

Peripheral T-cell lymphoma (PTCL) is a very aggressive and heterogeneous hematological malignancy and has no effective targeted therapy. The molecular pathogenesis of PTCL remains unknown. In this study, we chose the gene expression profile of GSE6338 from the Gene Expression Omnibus (GEO) database to identify hub genes and key pathways and explore possible molecular pathogenesis of PTCL by bioinformatic analysis. Differentially expressed genes (DEGs) between PTCL and normal T cells were selected using GEO2R tool. Gene ontology (GO) analysis and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway analysis were performed using Database for Annotation, Visualization and Integrated Discovery (DAVID). Moreover, the Search Tool for the Retrieval of Interacting Genes (STRING) and Molecular Complex Detection (MCODE) were utilized to construct protein-protein interaction (PPI) network and perform module analysis of these DEGs. A total of 518 DEGs were identified, including 413 down-regulated and 105 up-regulated genes. The down-regulated genes were enriched in osteoclast differentiation, Chagas disease and mitogen-activated protein kinase (MAPK) signaling pathway. The up-regulated genes were mainly associated with extracellular matrix (ECM)-receptor interaction, focal adhesion and pertussis. Four important modules were detected from the PPI network by using MCODE software. Fifteen hub genes with a high degree of connectivity were selected. Our study identified DEGs, hub genes and pathways associated with PTCL by bioinformatic analysis. Results provide a basis for further study on the pathogenesis of PTCL.

It is recognized that cystatin C is an effective marker for monitoring the glomerular filtration rate and the clinical diagnosis of various diseases. In this study, a novel immunochromatographic method has been established to achieve quantitative detection of serum cystatin C. Unlike conventional and traditional gold immune chromatographic assays (GICAs), a uniform layer of a fluorescent film was added to the solid phase, which has been designated as the background fluorescence quenching immune chromatographic assay (bFQICA). Under the optimized conditions, there was a good correlation between the fluorescence signal ratio (F1/F2) of the background fluorescence (F1) to the fluorescence signal at the detection line (F2) for cystatin C at concentrations from 0.0 ng/mL to 100 ng/mL with a correlation coefficient of 0.9977. The detection limit was 0.69 ng/mL, and the recovery values were 87.9-105%. The differences between the intra- and interbatch precision were less than 15% in three batches. In addition, after 120 serum samples were detected, there were no significant differences in the results obtained by bFQICA and immunoturbidimetry (t = 0.963, p = 0.338 > 0.05). This work demonstrates that bFQICA is a simple, sensitive, and accurate approach for the determination of serum cystatin C, providing a new approach for the clinical diagnosis of cystatin C.

Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease with unclear pathogenesis. DLBCL accounts for 30%–35% of all non-Hodgkin lymphomas (NHLs) and is an aggressive subtype of mature B-cell neoplasm. At present, half of DLBCL cases can be cured, although one-third of patients experience recurrence after treatment and enter advanced tumor stage. This study aimed to investigate the differentially expressed proteins in activated B-cell-like-DLBCL (ABC-DLBCL) through quantitative proteomics (iTRAQ). Seven ABC-DLBCL experimental samples and eight control samples (reactive hyperplasia of the lymph node) were obtained from fresh tissues. The exclusion criteria were expressed as follows: (1) patients with other lymphoid diseases; and (2) patients undergoing chemical treatment. A total of 5974 proteins were identified. P value < 0.05 and multiple expressions were more than 1.2-fold. A total of 131 upregulated and 204 downregulated differentially expressed proteins were identified. Gene ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway analysis were performed. Protein–protein interaction (PPI) network analysis was conducted. The expression levels of HSP90AB1, GNA13, LAMB2, LAMA5, YWHAZ, and IKBKB were evaluated through PRM and TCGA to validate the accuracy of iTRAQ and liquid chromatography–tandem mass spectrometry results. Results of differential multiple and t-test showed differences in the expression levels of six target proteins between the control and experimental groups. To the best of our knowledge, the present study is the first to identify proteins associated with ABC-DLBCL using iTRAQ technology. Our results provide new insights into the pathogenesis of ABC-DLBCL. The combination of ABC-DLBCL-associated signaling pathway proteins and targeted therapy to reverse drug resistance is of great significance in improving the comprehensive treatment of lymphoma and reducing mortality of affected individuals. The feasibility of the present study is limited due to the number of samples, and future studies are required to determine the function of proteins in ABC-DLBCL development.

Background fluorescence immunochromatography assay(bFQICA) has been established, which has the advantages of simple operation, high sensitivity, strong anti-interference and rapid quantitative detection of C-reactive protein(CRP). This method of using double antibody sandwich method principle, the appropriate concentration of capture antibody and tracer antibodies are respectively fixed on the test strip and the micropore. During chromatography, antibodies were captured, while tracer antibodies and samples were inserted to form a sandwich structure of antibody antigen antibody. The method has a good correlation with fluorescence signal values(F-1/F-2) in the range of 0.0-100.0 ng/mL concentration in CRP, the minimum detection limit was 0.0939 ng/mL, and the recovery rate was 87.69%-111.0%, three batches of reagents and inter and intra assay relative standard deviation was less than 15%, and procalcitonin(PCT, 20.0 ng/mL), serum amyloid A like protein(SAA, 10.0 mu g/mL) had not cross reaction with this method and the immune turbidimetric method for the simultaneous determination of 41 serum samples, the detection results had a good correlation(r=0.9585, P<0.01), there was no significant difference between the two methods(P>0.05). This method has high sensitivity, simple operation, and can be used for rapid and accurate quantitative detection of CRP.

Summary Background To establish a fast and simple quantitative method for detection of heart-type fatty acid-binding protein (H-FABP) in serum based on a background fluorescence quenching immunochromatographic assay. Methods A detection card based on the double-antibody sandwich double-antibody method with background fluorescence quenching was developed for quantitative measurement of H-FABP in serum. The optimal concentrations of control for coating the test and control lines were determined as well as the concentrations of gold-labeled antibodies used in preparing the detection system. The detection method for H-FABP in serum was established and validated using real-world clinical samples. Results The optimal concentrations of labeling antibody and coating antibody were 5.0 μg/mL and 1.0 mg/mL, respectively. The test card had a sensitivity of 1.15 ng/mL over a linear concentration range of 0–100 ng/mL. Based on three batches prepared for testing the card, the relative standard deviation (RSD) within batches was less than 15% without a significant difference (P=0.942). The detection method was tested against common interfering substances in serum, such as bilirubin, triglyceride and serum anticoagulants ethylenediamine tetraacetic acid (EDTA), heparin, and sodium citrate, and no significant cross-reaction was detected. The test method was further validated with 50 clinical serum samples, and the test results were comparable with standard reference detection methods with good correlation (R=0.95). Conclusion Our study presents a new method with strong specificity and sensitivity for the detection of H-FABP in serum, which could promote H-FABP detection in a broad range of applications.

The interaction between alliin and bovine serum albumin(BSA) and human serum albumin(HSA) was studied under a pH7. 4 Tris-HCl buffer system with multiple spectroscopic techniques. The study took 280 nm as the excitation wavelength to scan the range of 300 similar to 500 nm of fluorescence emission spectrum by using fluorescence spectrophotometer while setting wavelength differential as Delta lambda=-15 nm and Delta lambda=60 nm to scan synchronous fluorescence spectra. The fluorescence quenching data were analyzed by applying Stern-Volmer equation and Line weaver-Burk equation. It is shown that Alliin was effective in quenching BSA and HSA fluorescence. The binding constant obtained of alliin and BSA was 9. 81 x 10(2) L . mol(-1) at 298 K and 6. 15 x 10(2) L . mol(-1) at 310 K. The binding constant obtained of alliin and HSA was 2. 21 x 10(2) L . mol(-1) at 298 K and 6. 84 x 10(2) L . mol(-1) at 310 K. The thermodynamic parameters, Delta H calculated by using Gibbs-Helmholtz Equation of the reaction of alliin and BSA is -29. 9 kJ . mol(-1) and AS is 43. 0 J . morl . K-1 and AG is 17. 1 kJ . mol(-1) at 298 K. Delta H of the reaction of alliin and HSA is 72. 3 kJ . mol(-1) and AS is 288 J . mol(-1) . K-1 and AG is 13. 4 kJ . mol(-1) at 298 K. The main types of binding force of Alliin-BSA were hydrophobic interaction and the sorts of binding force of Alliin-HSA were electrostatic action. The results of synchronous fluorescence spectra showed that alliin has main effects on Tyr of BSA and both Tyr and Trp of HSA. The experimental results have provided the certain theory basis of the mechanism of interaction between alliin and biological small molecules.