1.Coupling of an Au@AgPt nanozyme array with an micrococcal nuclease-specific responsiveness strategy for colorimetric/SERS sensing of Staphylococcus aureus in patients with sepsis.
Xueqin HUANG ; Yingqi YANG ; Hanlin ZHOU ; Liping HU ; Annan YANG ; Hua JIN ; Biying ZHENG ; Jiang PI ; Jun XU ; Pinghua SUN ; Huai-Hong CAI ; Xujing LIANG ; Bin PAN ; Junxia ZHENG ; Haibo ZHOU
Journal of Pharmaceutical Analysis 2025;15(2):101085-101085
Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections. Herein, we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering (SERS) and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease (MNase) in serum samples. The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) molecules to SERS-enhanced oxidized TMB (oxTMB), accompanied by the color change from colorless to blue. In the presence of S. aureus, the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads (MBs) to release alkaline phosphatase (ALP), which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away. Using this "on-to-off" triggering strategy, the target S. aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode. Meanwhile, the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis (n = 7) and healthy participants (n = 3), as well as monitored the prognostic progression of the disease (n = 2). Overall, benefiting from highly active and dense "hot spot" substrate, MNase-mediated cascade response strategy, and colorimetric/SERS dual-signal output, this methodology will offer a promising avenue for the early diagnosis of S. aureus infection.
2.Research progress on the immune regulatory effect of myeloid derived suppressor cells on parasitic infections
Chinese Journal of Endemiology 2024;43(11):937-941
Myeloid derived suppressor cells (MDSCs) are a type of immature, differentiated, heterogeneous cells derived from the bone marrow. MDSC plays a dual role in parasitic infections. On the one hand, the cells can induce helper T cell 1/2 (Th1/Th2) immune responses by inducing host secretion of interferon-γ (IFN-γ), interleukin (IL)-4, and IL-13. On the other hand, they can also secrete IL-10, IL-17 and transforming growth factor-β (TGF-β) to induce regulatory T cells (Treg)/Th17 immune responses, thereby exerting immune protection or immune escape. This article provides a review of the research progress on the immune regulation of host T cells by MDSC in parasitic infections.
3.Research progress of LRRC15 protein in the medical field
Shunfu YU ; Qianqian MU ; Xiaoqing SUN ; Biying ZHOU
Chinese Journal of Endemiology 2024;43(11):942-946
The leucine-rich repeat (LRR) containing 15 (LRRC15) protein is one of the members of the LRR superfamily. In the medical field, LRRC15 protein not only participates in parasitic, viral and other pathogen infections, but also plays a role in anti-tumor and rheumatoid arthritis treatment. It also regulates cell autophagy, stem cell ossification, and changes in hair follicle dermal papilla cell characteristics. This article reviews the research progress of LRRC15 protein in the medical field.
4.Prokaryotic expression and polyclonal antibody preparation of LRRC15 protein, an excretory-secretory antigen of Cysticercus cellulosae
Qianqian WANG ; Shimin WANG ; Qianfei WANG ; Fengling YUAN ; Wei HE ; Lizhu LI ; Biying ZHOU
Chinese Journal of Endemiology 2023;42(9):704-709
Objective:To construct a recombinant plasmid pET30a-leucine-rich repeat (LRR) containing 15 (LRRC15) of Taenia solium, prokaryotically express and purify the LRRC15 recombinant protein, and prepare a rabbit polyclonal antibody. Methods:The LRRC15 protein encoding gene of Taenia solium was obtained by whole gene synthesis; it was cloned into pET30a vector, and the recombinant plasmid pET30a-LRRC15 was constructed and identified by double-enzyme PCR; the recombinant plasmid was transformed into competent cells of Escherichia coli BL21 (DE3), and the recombinant protein LRRC15 was induced to express by isopropyl-beta-D-thiogalactopyranoside (IPTG), the expression product was analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); the LRRC15 recombinant protein was purified by Ni-IDA affinity columns, the purified recombinant protein was analyzed and identified by SDS-PAGE, and the specificity of the purified recombinant protein was identified by Western blot (WB); the New Zealand rabbits were immunized with purified LRRC15 recombinant protein to prepare polyclonal antibodies against LRRC15, and the potency of the purified polyclonal antibody was determined by indirect enzyme-linked immunosorbent assay (ELISA). Results:After PCR identification, a band with a length of 1 506 bp was amplified, which was consistent with the LRRC15 gene; after SDS-PAGE and WB identification, the LRRC15 target protein with a relative molecular mass ( Mr) of about 55.36 × 10 3 was obtained; after immunizing New Zealand rabbits with purified LRRC15 recombinant protein, a polyclonal antibody against LRRC15 was obtained, and its potency was 1∶1 587 200. Conclusion:The recombinant plasmid pET30a-LRRC15 is successfully constructed, the LRRC15 recombinant protein of Taenia solium is prepared, and a high purity and high potency rabbit anti polyclonal antibody against LRRC15 recombinant protein is obtained.
5.Research progress on the influence of TGF-β/Smad signaling pathway on host T cell immune responses in parasite infection
Xiaoqing SUN ; Wei HE ; Biying ZHOU
Chinese Journal of Endemiology 2023;42(9):758-762
Parasitic infection are an important global public health issue, and parasites can modulate and evade host immune attacks through various ways. In recent years, studies have shown that in the process of parasite infection, transforming growth factor-β (TGF-β)/Smad signaling pathway participates in host T cell immune response, induce T cells to proliferate and differentiate towards regulatory cells (Treg) and helper T cells (Th), regulate Treg/Th17 cell balance, inhibit Th1 cell proliferation and differentiation, and play an important role in the occurrence and development of parasitic diseases and their interactions with the host. This article reviews the research progress of T cell immune responses mediated by TGF-β/Smad signaling pathway in the process of parasite infection.
6.RNA Methylome Reveals the m6A-mediated Regulation of Flavor Metabolites in Tea Leaves under Solar-withering.
Chen ZHU ; Shuting ZHANG ; Chengzhe ZHOU ; Caiyun TIAN ; Biying SHI ; Kai XU ; Linjie HUANG ; Yun SUN ; Yuling LIN ; Zhongxiong LAI ; Yuqiong GUO
Genomics, Proteomics & Bioinformatics 2023;21(4):769-787
The epitranscriptomic mark N6-methyladenosine (m6A), which is the predominant internal modification in RNA, is important for plant responses to diverse stresses. Multiple environmental stresses caused by the tea-withering process can greatly influence the accumulation of specialized metabolites and the formation of tea flavor. However, the effects of the m6A-mediated regulatory mechanism on flavor-related metabolic pathways in tea leaves remain relatively uncharacterized. We performed an integrated RNA methylome and transcriptome analysis to explore the m6A-mediated regulatory mechanism and its effects on flavonoid and terpenoid metabolism in tea (Camellia sinensis) leaves under solar-withering conditions. Dynamic changes in global m6A level in tea leaves were mainly controlled by two m6A erasers (CsALKBH4A and CsALKBH4B) during solar-withering treatments. Differentially methylated peak-associated genes following solar-withering treatments with different shading rates were assigned to terpenoid biosynthesis and spliceosome pathways. Further analyses indicated that CsALKBH4-driven RNA demethylation can directly affect the accumulation of volatile terpenoids by mediating the stability and abundance of terpenoid biosynthesis-related transcripts and also indirectly influence the flavonoid, catechin, and theaflavin contents by triggering alternative splicing-mediated regulation. Our findings revealed a novel layer of epitranscriptomic gene regulation in tea flavor-related metabolic pathways and established a link between the m6A-mediated regulatory mechanism and the formation of tea flavor under solar-withering conditions.
RNA/metabolism*
;
Epigenome
;
Plant Proteins/metabolism*
;
Plant Leaves/metabolism*
;
Camellia sinensis/metabolism*
;
Flavonoids
;
Terpenes/metabolism*
;
Tea/metabolism*
;
Gene Expression Regulation, Plant
7.Transcriptome analysis reveals the role of withering treatment in flavor formation of oolong tea (Camellia sinensis).
Chen ZHU ; Shuting ZHANG ; Chengzhe ZHOU ; Biying SHI ; Linjie HUANG ; Yuling LIN ; Zhongxiong LAI ; Yuqiong GUO
Chinese Journal of Biotechnology 2022;38(1):303-327
Oolong tea is a semi-fermented tea with strong flavor, which is widely favored by consumers because of its floral and fruity aroma as well as fresh and mellow taste. During the processing of oolong tea, withering is the first indispensable process for improving flavor formation. However, the molecular mechanism that affects the flavor formation of oolong tea during withering remains unclear. Transcriptome sequencing was used to analyze the difference among the fresh leaves, indoor-withered leaves and solar-withered leaves of oolong tea. A total of 10 793 differentially expressed genes were identified from the three samples. KEGG enrichment analysis showed that the differentially expressed genes were mainly involved in flavonoid synthesis, terpenoid synthesis, plant hormone signal transduction and spliceosome pathways. Subsequently, twelve differentially expressed genes and four differential splicing genes were identified from the four enrichment pathways for fluorescence quantitative PCR analysis. The results showed that the expression patterns of the selected genes during withering were consistent with the results in the transcriptome datasets. Further analysis revealed that the transcriptional inhibition of flavonoid biosynthesis-related genes, the transcriptional enhancement of terpenoid biosynthesis-related genes, as well as the jasmonic acid signal transduction and the alternative splicing mechanism jointly contributed to the flavor formation of high floral and fruity aroma and low bitterness in solar-withered leaves. The results may facilitate better understanding the molecular mechanisms of solar-withering treatment in flavor formation of oolong tea.
Camellia sinensis/genetics*
;
Gene Expression Profiling
;
Plant Leaves
;
Plant Proteins/metabolism*
;
Taste
;
Tea
;
Transcriptome/genetics*
8.Research progress of recombinant thioredoxin peroxidase of important human parasites involved in immunoregulation, immunodiagnosis and immunoprophylaxis
Wei HE ; Bo LUO ; Biying ZHOU
Chinese Journal of Endemiology 2022;41(10):856-860
Thioredoxin peroxidase (TPx) belongs to the superfamily of peroxiredoxins, which is widely expressed in various growth and development stages of parasites and their excretory secretions. On the one hand, recombinant TPx protein can participate in host immunoregulation; on the other hand, recombinant TPx protein has high sensitivity and specificity as a diagnostic antigen, and can be used for immunodiagnosis of parasitic diseases; in addition, it can also be used as a candidate vaccine molecule for the immunoprophylaxis of parasitic diseases. This paper reviews the research progress on host immunoregulation, immunodiagnosis and immunoprophylaxis by recombinant TPx protein of important human parasites.
9.Research on the application of flipped classroom based on MOOC in human parasitology teaching
Chinese Journal of Medical Education Research 2021;20(1):19-21
In order to improve the quality of teaching, the teaching and research section of parasitology explores the teaching effects of human parasitology based on independent learning. Taking the clinical medicine specialty (excellent doctor class) as an example, in order to overcome the problems that the teaching content brought by traditional infusion teaching is difficult to meet the needs of modern teaching and students have low interest in learning. On the basis of optimizing teaching content and teaching methods, some courses are adopted flipped classroom teaching model based on the MOOC videos learning before class, emphasizing student-centered and self-directed learning, focusing on independent learning ability and critical thinking training, and introducing modern information technology and improving teaching effectiveness. The teaching effect shows that the student-centered teaching thought can be better reflected, students' ability of self-learning has been enhanced, and their sense of learning achievement has been strengthened, in favor of cultivating their ability to analyze and solve problems, as well as lifelong learning ability and teamwork ability.
10.Research progress of immunoregulation of T lymphocytes in cysticercosis
Chinese Journal of Endemiology 2021;40(2):164-168
Taenia solium is one of the main human parasites. Both larvae and adults can cause diseases, among which neurocysticercosis caused by Cysticercus is the most serious disease. The immune response induced by Cysticercus infection and its regulatory mechanism have been the key problems in the prevention and treatment of Cysticercus infection. This review reports the progress of T lymphocytes-mediated immune responses and immunoregulation in cysticercosis, which is expected to provide a theoretical basis for the immunopathogenesis and prevention and treatment of cysticercosis.

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