(1. The First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan 250014, China; 2. College of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China; 3. Weifang Hospital of Traditional Chinese Medicine, Weifang 261000, China; 4. Yanqing Hospital of Beijing Chinese Medicine Hospital, Beijing 102100, China; 5. Affiliated Hospital of Shandong University of Chinese Medicine, Jinan 250011, China)
Abstract: Islet β cells are the exclusive cellular source of insulin synthesis and secretion, playing a crucial role in regulating blood glucose homeostasis. The impairment of islet β cells due to sustained anaerobic glycolysis pressure represents a significant factor contributing to insulin deficiency or resistance in patients with type 2 diabetes mellitus (T2DM). Glucokinase (GK), serving as the principal rate-limiting step in glucose transport, exhibits close association with insulin secretion. Hypoxia-inducible factor 1α subunit (HIF-1α) and 6-phosphofructo-2-kinase (PFKFB3) act as pivotal regulatory factors within glycolysis. The HIF-1α/PFKFB3 signaling pathway participates in intricate pathophysiological processes involving islet β cell function and maintenance under high-glucose conditions. This review elucidates the mechanisms by which the HIF-1α/PFKFB3 signaling pathway mediates GK induction through anaerobic glycolysis, mitochondrial network adaptive fragmentation, oxidative stress, and dedifferentiation leading to pancreatic islet β cell dysfunction in T2DM, thereby offering novel insights for clinical prevention and treatment strategies targeting T2DM.
Key words: type 2 diabetes; glucokinase; HIF-1α/PFKFB3 signaling pathway; mitochondria; dedifferentiation
（Acta Laser Biology Sinica, 2024, 33(6): 512-522）

Abstract: Synthetic biology can fully promote the efficient bio-manufacturing of target natural products in the aspects of efficient chassis cell construction, active natural product mining and metabolic pathway optimization. However, in the study of synthetic biology of Saccharopolyspora spinosa, due to the limited understanding of the important functional genes regulating spinosyn biosynthesis and their action mechanisms, it is difficult to construct efficient cell factories to significantly increase the spinosyn production through the “design-build-test-learn” strategy. In order to solve this problem, an excellent chassis strain CW-12 with high spinosyn production, fast growth rate and strong extracellular glucose uptake ability was obtained by artificial mutagenesis, and comparative proteomic analysis was performed. The results showed that the enhancement of metabolic pathways related to intracellular carbon metabolism, fatty acid metabolism, amino acid biosynthesis, and TCA cycle was an important reason for the improvement of spinosyn biosynthesis of mutant CW-12, and 668 up-regulated proteins were screened. Subsequently, we selected 3-hydroxyacyl-CoA dehydrogenase (SS_2202) and acetyl-CoA acetyltransferase (SS_2203) to analyze their action mechanisms, confirming that their overexpression can effectively promote the spinosyn biosynthesis. This study has certain guiding significance on how to explore the important functional genes regulating spinosyn biosynthesis and the analysis of their action mechanisms, and lays an important foundation for the subsequent construction of spinosyn efficient biosynthesis cell factories through synthetic biology strategies.
Key word: Saccharopolyspora spinosa; spinosyn; synthetic biology; comparative proteomic analysis; gene editing
（Acta Laser Biology Sinica, 2024, 33(5): 450-460）

Abstract: Water pollution is one of the most serious environment problems that affect people′s life quality and sustainable economic development worldwide. The degradation capability of microorganisms endows them advantages in environmental pollutants elimination and water environments repairment, with the utilization of diverse bioremediation technologies. However, due to the complex situation of water pollution environment, traditional microbiological treatment methodology is restricted and limited. With the engineering principle Design-Build-Test-Learn in synthetic biology, the designed and modified synthetic microorganisms with efficient and broad-spectrum degradation ability for pollutants has attracted the researchers′ attention. The synthetic microorganisms could not only meet the wastewater treatment requirements within complex pollutants, but also could achieve the green, efficient circular treatment goal. Therefore, design and construction of synthetic microorganisms for water pollution control has becoming one of the future development and application direction of synthetic biology. Herein, we introduced the current situation of water pollution, the treatment technologies classically utilized, and the synthetic microorganisms′ application in water pollution control. This review focuses on different types of microbial chassis in synthetic biology and the merits of application of synthetic microorganisms in water pollution control. This review will facilitate the performance optimization of pollutant degradation and wastewater treatment via synthetic biology and provide reference for the efficient treatment of wastewater environment.
Key words: synthetic microorganisms; microbial chassis; water pollution; bioremediation technology; environmental governance
（Acta Laser Biology Sinica, 2024, 33(5): 418-431）

Abstract: The special connection approaches of linear ubiquitin chain have been elucidated in recent years. Currently, the types of commercial linear ubiquitin chain specific antibodies are limited and their specificity still needs to be improved. RHD1 is a papain-like deubiquitinating enzyme that specifically recognizes and cleaves linear ubiquitin chains. The mutation at C13 in its enzymatic activity site results in the loss of deubiquitination function for the RHD1. Meanwhile, the RHD1 with this mutation still exhibits low binding activity towards linear ubiquitin chain. On the basis of this mutant, we optimized the mutations at interaction interface between RHD1 and Di-Ub (M1-linked) in order to enhance the specific binding activity of RHD1. In this study, we utilized the structural model of RHD1 and Di-Ub (M1-linked) complex to analyze the physical and chemical properties of all amino acids at the interaction interface and subsequently designed a RHD1 mutant library. The site-mutations in RHD1 variant polypeptides were generated by using overlap extension PCR method, and all recombinant variant proteins were expressed by using E. coli BL21 (DE3) strain. By employing the enzyme-linked immune sorbent assay (ELISA ) method for screening, we successfully identified a RHD1 variant harboring C13A and A92T mutations, which showed significantly increased affinity to Tetra-Ub (M1-linked). This RHD1 variant holds great potential as a valuable tool for studying linear ubiquitin chains, and exhibits promising application in clinical diagnostics and the development of related therapeutic agents.
Key words: RHD1 deubiquitinating enzymes; Tetra-Ub (M1-linked); site-specific mutation; prokaryotic expression; ELISA
（Acta Laser Biology Sinica, 2024, 33(5): 439-449）

Abstract: L-methionine is an essential amino acid with a wide range of applications in food, feed, cosmetics and pharmaceuticals. Currently, L-methionine is the only essential amino acid that cannot be industrially produced by microbial fermentation. In recent years, the use of metabolic engineering to enhance the yield of L-methionine has received widespread attention from researchers at home and abroad. In this paper, the biosynthetic pathway and regulatory mechanism of L-methionine in Corynebacterium glutamicum and Escherichia coli are analysed; three key steps (i.e., acylation, sulfurylation, and methylation) in the biosynthesis of methionine from hyper-serine are highlighted, and a further outlook on the biosynthesis of L-methionine is proposed with a view to providing guidance for the industrial production of L-methionine.
Key words: L-methionine; biosynthesis; regulatory mechanism
（Acta Laser Biology Sinica, 2024, 33(5): 408-417）

Abstract: Glutamate transporter GluA, is a class of ATP-binding proteins that can affect intracellular carbon metabolism, nitrogen metabolism and secondary metabolism by regulating the active transport of glutamate, belonging to the ABC transport system. To study the effects of GluA on the strain growth development and butenyl-spinosyn biosynthesis in Saccharopolyspora pogona, gluA-1 overexpression and deletion strains were constructed by genetic engineering technology. Phenotypic analysis showed that the butenyl-spinosyn production increased by 4.2 times, the bacterial growth logarithmic phase was prolonged, the maximum bacterial density and acetyl-CoA content significantly increased, whereas the mycelium morphology was not significantly changed in overexpressed strain S. pogona::gluA-1. The butenyl-spinosyn production decreased by 37%, the strain growth was inhibited obviously, the acetyl-CoA content significantly decreased, and the mycelium was shortened and thickened, branching was reduced in deletion strain S. pogona-ΔgluA-1. These results indicated that GluA could promote the growth development and the butenyl-spinosyn biosynthesis in S. pogona, which laid an important foundation for the study of GluA's role in the secondary metabolism of Streptomyces.
Key word: Saccharopolyspora pogona; butenyl-spinosyn; glutamate transporter protein; biology synthesis; gene editing
（Acta Laser Biology Sinica, 2024, 33(5): 470-480）

Abstract: Spinosad, a secondary metabolite produced by Saccharopolyspora spinosa, has garnered significant attention globally due to its environmentally friendly and broad-spectrum insecticidal properties. The unique structure of spinosad and its analog, butenyl-spinosyn, underlies their distinctive mode of action. To meet the demands of large-scale industrial production, research teams worldwide have been striving to enhance the yields of spinosad and butenyl-spinosyn, with remarkable achievements accomplished through the application of synthetic biology techniques. This review summarizes the research conducted domestically and internationally on the regulation of metabolic pathways at various levels using synthetic biology techniques to improve the production of spinosad and butenyl-spinosyn. It focuses on four key aspects: chassis cell modification and optimization, multi-omics analysis, genetic circuit modification, and metabolic network regulation. Furthermore, it provides an outlook on future research directions in this field, and provides a new research strategy to promote the biosynthesis of spinosad by synthetic biology technology.
Key words: spinosad; Saccharopolyspora spinosa; synthetic biology; gene editing; metabolic pathway regulation
（Acta Laser Biology Sinica, 2024, 33(5): 400-407）

Abstract: Edeine, synthesized by Brevibacillus brevis X23, is a linear peptide antibiotic not encoded by ribosomal RNA, renowned for its extensive range of antibacterial effectiveness. Transcriptome sequencing revealed an inverse expression pattern between the two-component regulatory factors yvrG/yvrH and the edeine biosynthesis gene cluster (ede BGC), suggesting the involvement of YvrH/YvrG in the transcriptional regulation of ede BGC. This study utilized Red/ET homologous recombination technology to construct X23 mutant strains that yvrG/yvrH lacks, complemented strains, and overexpressing strains, and investigated the impact of YvrG/YvrH on edeine biosynthesis through phenotype analysis and quantitative real-time PCR (RT-qPCR) techniques. The results revealed that knocking out yvrG/yvrH significantly reduced the antibacterial activity of the strains, the expression of ede BGC decreased, the synthesis of edeine was inhibited, and the yield of edeine was reduced by 46.75%. In summary, YvrG/YvrH act as positive regulatory factors in the edeine biosynthesis. This study provides gene regulatory elements for the metabolic engineering of edeine.
Key words: Brevibacillus brevis; edeine; two-component regulatory factors; bacteriostatic activity; biosynthesis
（Acta Laser Biology Sinica, 2024, 33(5): 461-469）

Abstract: One-carbon compounds (C1), including methane, formate, methanol, carbon monoxide and carbon dioxide, have been considered to be the ideal raw materials for the third generation of biorefining, due to their abundance and low cost. Microbes that can naturally utilize C1 have been widely studied with low carbon conversion rate and industrial difficulty. The transformation and heterologous expression of C1 utilization pathway described as synthetic one-carbon bio-utilization system can help overcome the obstacles of non-model organisms, which will reduce the dependence on traditional fossil fuels. This review briefly summarized the types, sources and natural pathways of C1. We mainly focused on the progress of C1 utilization in model organisms and emphasized the importance of synthetic C1 bio-utilization systems for improving carbon conversion rate and developing new application. In addition, we discussed the challenges and perspectives of the biotransformation of C1. This paper provided a reference for the construction of synthetic C1 utilization microorganisms and improving the efficiency of heterologous C1 metabolic pathway, which is of great significance for realizing green and efficient development of sustainable resources.
Key words: one-carbon compounds; biotransformation; carbon dioxide; synthetic one-carbon bio-utilization system; carbon conversion rate
（Acta Laser Biology Sinica, 2024, 33(5): 385-399）

Abstract: Methyltransferases (MTs) are a class of enzymes that are ubiquitous in biological organisms, usually using S-adenosylmethionine as a methyl donor to catalyze the methylation reaction of the substrate. The heterologous expression of MTs in microorganisms has made great progress in realizing the biosynthesis of some important natural products. MTs can be used in microorganisms to synthesize important natural products such as phenylpropanoids, fragrance compounds, hormones and antibiotics. MTs have also been widely used in many fields such as medicine, chemical industry and energy, showing great application value and broad application prospects. In this review, we summarize the classification, function and application of natural product methyltransferases, in order to provide theoretical guidance for the efficient artificial biosynthesis of highly active non-ribosomal peptide synthetase (RXPs) peptides.
Key words: methyltransferases; S-adenosylmethionine; methylation; natural products; biosynthesis
（Acta Laser Biology Sinica, 2024, 33(5): 432-438）

(State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin 300350, China)
Abstract: The high porosity and customizable structure of metal-organic frameworks (MOFs) allow them to bind a wide range of biomolecules and have great potential as biosensors. MOFs can exhibit luminescence from metal nodes, ligands or introduced guests, which provides an excellent fluorescence response for sensing. However, single-signal emitting MOFs are susceptible to interference from concentration, environment, and excitation intensity, resulting in poor accuracy. Dual-fluorescence-emitting MOFs biosensors overcome these shortcomings and need to be fully and accurately characterized. Fluorescence resonance energy transfer (FRET) is the process in which the internal energy of fluorescent molecules is transferred from the donor to the acceptor, and FRET detection of dual-emission MOFs based on laser scanning confocal microscopy can achieve visual imaging and accurate efficiency analysis, and has become an effective evaluation method for dual-emission MOFs. In this paper, the technical process and conditions of FRET for the detection of dual-emission MOFs are described, and combined with the research examples of Alzheimer's disease and COVID-19, the application progress of MOFs fluorescent biosensors in terms of design concept, binding mode, and action efficiency is demonstrated. The results confirmed that this technique has high detection sensitivity and specificity, which provides a new idea for probe development and evaluation.
Key words: fluorescence resonance energy transfer; metal-organic frameworks; laser confocal microscopy; fluorescent
biosensors; double fluorescence emission
（Acta Laser Biology Sinica, 2024, 33(6): 481-488）

(1. Institute of Subtropical Agriculture, Chinese Academy of Sciences, Key Laboratory for Agro-ecological Processes in Subtropical Region, National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production, Hunan Provincial Key Laboratory of Animal Nutritional Physiology and Metabolic Process, Changsha 410125, China; 2. University of Chinese Academy of Sciences, Beijing 100049, China)
Abstract: The present study aimed to evaluate the effect of Laminaria japonica extract (LJE) supplementation on lactation performance, plasma biochemistry, immunity, antioxidant status, rumen fermentation parameters and rumen microbiota of dairy cows. The experiment was conducted in a one-way completely randomized design. 32 healthy lactating Holstein dairy cows with similar body weight, milk yield, parity and days in milk were selected and randomly assigned to 2 groups of 16 cows each. The cows in the control group (CON group) fed with a basal diet, and the other group (LJE group) received the CON diet supplemented with LJE at 30 g per cow per day. The entire trial includes a 12-day adaptation and a 30-day trial period. Compared with the CON group, the diet supplemented with LJE increased dry matter intake (DMI), milk yield, and milk protein content of dairy cows, whereas it had no significant effects on milk fat, lactose, somatic cell counts, and total solids, and it had no significant effects on the plasma biochemical indices. Moreover, the LJE group had significantly higher levels of immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in plasma, and had lower malondialdehyde content than those of the CON group. The total volatile fatty acid level increased in the rumen of dairy cows. The LJE diet increased the relative abundance of the phylum Firmicutes and the genera Succiniclasticum, Sphaerochaeta, Mucispirillum, and Akkermansia. In conclusion, adding LJE in feed can enhance the immunity of dairy cows, and reduce the level of lipid peroxidation in dairy cows, and improve the DMI, milk yield, and milk quality, which is of great significance to realize the healthy breeding of dairy cows.
Key words: Laminaria japonica extract; milk performance; immunity; rumen fermentation; rumen microbiota
（Acta Laser Biology Sinica, 2024, 33(6): 530-540）

Abstract: Taihu muskmelon, commonly known as “Laotaipo muskmelon” and “crisp muskmelon”, has a long history of cultivation. The skin of the muskmelon is white, the aroma is strong, the meat is crisp and sweet, and it is loved by consumers. However, there is a lack of research on the identification of the characteristics of Taihu muskmelon and the changes of metabolites during storage. The metabolites of Xiaobainiang, Yunaixiang 2 and Lüxiangtian were analyzed by ultra-high performance liquid chromatography-high resolution mass spectrometry. At the same time, the Xiaobainiang was stored at room temperature for 1 d and 8 d, and the metabolites of its flesh were analyzed. Using common muskmelon varieties in the market, Lüxiangtian and Yunaixiang 2, as controls, a total of 18 136 metabolites were identified in the flesh of the three muskmelon varieties, Xiaobainiang, through metabolomics analysis, which was higher than the control of 14 879 metabolites in Lüxiangtian and 15 943 metabolites in Yunaixiang 2. Among them, a total of 1 902 unique metabolites were identified in Xiaobainiang, which was higher than the 449 unique metabolites in the control group of Lüxiangtian and 542 unique metabolites in Yunaixiang 2. Analysis of the content of metabolites revealed that the flavonoid compound kaempferol-3-gentiobioside in Xiaobainiang was significantly higher than that in Lüxiangtian and Yunaixiang 2, which were 136 and 3 200 times higher, respectively. Meanwhile, the metabolites of Xiaobainiang muskmelon were measured after being stored at room temperature for 1 and 8 days, and principal component analysis (PCA)  was performed on the differential metabolites. The results showed that there were no significant differences in the differential metabolites. In conclusion, Xiaobainiang not only has a certain level of storage stability, but also contains a high amount of flavonoid compounds, particularly kaempferol-3-gentiobioside, which can serve as a characteristic marker for identifying Taihu fragrant melon Xiaobainiang. This study is of significant importance for the identification and promotion of Xiaobainiang.
Key words: Taihu muskmelon; metabolome; characteristics; storage stability
（Acta Laser Biology Sinica, 2025, 34(1): 077-083）

(Affiliated Hospital of Xuzhou Medical University, Xuzhou 221000, China)
Abstract: Acute pancreatitis（AP） is one of the most common gastrointestinal diseases requiring urgent hospitalization, and its incidence is on the rise. This study aims to evaluate whether empagliflozin (EMPA) can alleviate AP in rats by inhibiting apoptosis. Eighteen SPF-grade SD male rats were randomly divided into three groups using a random number table method: control group (CON group), AP group, and AP+EMPA group, with six rats in each group. After successful establishment of the rat models, blood and pancreatic samples were collected. The serum markers tested included amylase (AMY), glucose (GLU), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) levels. Western blot analysis was used to detect the expression levels of Caspase-3 and Bcl2 in the pancreatic tissue, while immunohistochemistry and immunofluorescence were used to assess Caspase-3 expression in the pancreatic tissue. Pancreatic tissues were paraffin-embedded for hematoxylin-eosin (HE) staining and pathological scoring. One-way ANOVA was employed for statistical analysis of the data. The study found that the AP group had elevated serum AMY, GLU, inflammatory cytokine levels, and pathological scores compared to the CON group. In the pancreatic tissue, the apoptosis-related molecule Caspase-3 was elevated, while the expression level of Bcl2 was reduced compared to the CON group (P<0.05). However, EMPA treatment could reverse these changes. In conclusion, EMPA effectively ameliorates the pathological changes in the pancreatic tissue in a rat model of AP, and this protective effect may be related to the inhibition of apoptosis. This article aims to explore the potential therapeutic effects and molecular mechanisms of EMPA in AP, providing new insights for the effective prevention and treatment of this disease.
Key words: acute pancreatitis; apoptosis; Caspase-3; empagliflozin; rat model
（Acta Laser Biology Sinica, 2024, 33(6): 550-556）

(1. Laboratory of Animal Nutrition and Human Health, Hunan Normal University, Changsha, 410081, China; 2. Chenzhou Science and Technology Innovation Service Center, Chenzhou 423000, China)
Abstract: The hoxc8a gene is a member of the Hox gene family, which is evolutionarily highly conserved. It has been studied that the hoxc8a gene is expressed in the swim bladder and pectoral fins of zebrafish, and that Hox genes can affect the formation and development of vertebrate body axes, whereas the mechanism of action of the hoxc8a gene is not clear.  In order to study the developmental biology functions and its molecular regulation mechanism of this gene, the gene will be knocked out using CRISPR/Cas9 gene editing technology. First, target sites as well as genotype detection primers were designed on exon 1 of the hoxc8a gene, and guide DNA (sgDNA) was obtained by PCR. Then the sgDNA was recovered and purified, and the purified sgDNA was used as a template to synthesize guide RNA (sgRNA) by in vitro transcription, and finally, RNA was purified. The sgRNA and Cas9 protein were mixed in a 1:1:0.8 ratio, and the mixture was injected into wild-type zebrafish embryos using microinjection technique, and the mutant F0 generation zebrafish were screened by genotyping. When the F0 generation zebrafish were sexually mature, they were crossed with wild-type zebrafish, and the offsprings were genotyped and screened to obtain the mutant heterozygotes of the F1, and the F1 generation of zebrafish with the protein code-shifting mutation were raised after Sanger sequencing, and continued to be bred to adult. The F1 zebrafish were crossed, and the offsprings were genotyped and screened to obtain F2 homozygotes. Finally, a calcium staining test was performed on mutant homozygous and wild-type zebrafish, and it was found that the spine development of the knockout mutant was not significantly different from that of the wild-type, suggesting that the hoxc8a gene does not affect the early spine development of zebrafish, whereas whether the gene affects the development of other tissues and organs remains to be further investigated. Thus, by constructing zebrafish hoxc8a knockout lines, thus we can provide a research basis for more in-depth studies of the hoxc8a gene.
Key words: zebrafish; hoxc8a; spinal development; gene knockout line; homozygous mutant
（Acta Laser Biology Sinica, 2024, 33(6): 557-566）

(1. Hunan Institute of Traffic Engineering, Hengyang 421000, China; 2. Xiangxing College, Hunan University of TCM, Xiangyin 410007, China; 3. College of Life Science, Hunan Normal University, Changsha 410081, China; 4. Department of Orthopedics, Xiangya Hospital, Central South University, Changsha 410008, China)
Abstract: Osteoporosis is the most common systemic and metabolic bone disease. The concentration of serum resistin (RETN) is associated with bone mineral density, but it is still unclear whether there is a causal relationship between RETN and bone density and fractures. This study used Mendelian randomization (MR) to analyze the causal relationship between RETN gene expression and bone mineral density and fractures, and then analyzed the differential expression of RETN gene in single-cell sequencing data of femeral neck fracture (FNF) and osteoarthritis (OA) samples. MR analysis showed that high expression of RETN was found to lead to decreased bone mineral density and increased risk of fractures. RETN expression was upregulated in the FNF group, and RETN was mainly expressed by neutrophils. The inflammatory chemokine CXCL8 related to RETN was significantly upregulated in the FNF group. This study suggested the role of RETN in the pathogenesis of osteoporosis, providing a basis for the diagnosis and treatment of osteoporosis.
Key words: osteoporosis; RETN; Mendelian randomization; single-cell RNA sequencing; bone mineral density
（Acta Laser Biology Sinica, 2024, 33(6): 541-549）

(College of Management, Gansu Agricultural University, Lanzhou 730070, China)
Abstract: This paper aims to explore the application of radio frequency identification (RFID) and laser scanning based internet of things (IoT) technology in clinical trials, and analyze its advantages in improving data accuracy, improving work efficiency, and ensuring data security. RFID technology realizes the automatic identification and collection of items and data through radio waves, which has the characteristics of no contact reading, high capacity data storage and real-time information update. Laser scanning technology uses a laser beam to scan a bar code or quick response (QR) code to obtain information, with high precision and high speed. Both techniques performed well for subject identity verification, sample management, and medication tracking. In specific applications, RFID and laser scanning technologies have greatly improved the transparent management and traceability of trial materials, ensuring the real-time collection and monitoring of subject data, and safe distribution and use of drugs. These techniques reduce human error and improve data accuracy and work efficiency by automating and transmitting information in real time. Despite the challenges of technology costs, privacy protection, and standardization, these issues are expected to be gradually resolved with technological progress and innovation. In the future, RFID and laser scanning technology will be further integrated with emerging technologies such as artificial intelligence and blockchain to promote the digital and refined management of clinical trials. The research and analysis in this paper provide a reference for the digital transformation of clinical trials, hoping to promote the wide application of RFID and laser scanning technology in this field, and contribute to the innovation and progress of medical research.
Key words: radio frequency identification; laser scanning; automatic recognition; internet of things technology; clinical trial
（Acta Laser Biology Sinica, 2024, 33(6): 505-511）

(1. Hunan Polytechnic of Environment and Biology, Hengyang 421005, China; 2. Hunan Institute of Microbiology, Changsha 410009, China)
Abstract: The study of rumen microbiota is of great significance for understanding the health, growth and development, as well as digestion and absorption processes of ruminants. In recent years, with the development of high-throughput sequencing technology and the rise of microbiome research, significant progress has been made in the study of rumen microbiota in ruminants. This article mainly reviews the composition and influencing factors of rumen microbiota in ruminants, as well as the technical methods for studying rumen microbiota, including 16S rRNA gene sequencing, functional genomics analysis, metabolomics research, and fluorescence in situ hybridization (FISH). Meanwhile, the application and development prospects of these methods in the study of rumen microbiota in ruminants will also be explored, aiming to provide reference for the structural and functional analysis of rumen microbiota.
Key words: ruminant animals; rumen microbiota; high throughput sequencing; microbial community; multiomics joint analysis
（Acta Laser Biology Sinica, 2024, 33(6): 496-504）

（Department of Stomatology, Hebei Petro China Central Hospital, Langfang 065000, China）
Abstract: To investigate the effects of luteic acid on inflammation and apoptosis of human periodontal stem cells induced by lipopolysaccharide and the regulation of Wnt/β-catenin signaling pathway, hPDLSCs were cultured in vitro, which was divided into two steps: pre-experiment and formal experiment. The pre-experiment was divided into control group (no intervention), lipopolysaccharide group (10 μg/mL lipopolysaccharide), low/medium/high concentration luboflavic acid group (10 μg/mL lipopolysaccharide +0.5, 1.0, 2.0 μmol/L luboflavic acid). The formal experiment was divided into control group, lipopolypaccharide group, luteic acid group (10 μg/mL lipopolypaccharide +2.0 μmol/L luteic acid), inhibitor group (10 μg/mL lipopolypaccharide +2 μmol/L Wnt/β-catenin signaling pathway inhibitor XAV939), luteic acid + inhibitor group (10 μg/mL lipopolysaccharide +2.0 μmol/L luteic acid +2 μmol/L XAV939) and luteic acid + activator group (10 μg/mL lipopolysaccharide +2.0 μmol/L luteic acid +20 μmol/L Wnt/β-catenin signaling pathway activator SKL2001), they were treated for 24 h. Cell viability was measured by CCK-8. The expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were detected by enzyme-linked immunosorbent assay (ELISA). The apoptosis rate was detected by Hoechst 33258 staining. The expression levels of B lymphoblastoma-2 (Bcl-2), cysteine aspartate protease-3 (Caspase-3) and Wnt/β-catenin signaling pathway were detected by Western blot (Wb). In the preliminary experiment, the cell viability of LPS group decreased compared with control group (P<0.05), and the expression levels of inflammatory factors TNF-α, IL-1β and IL-6 increased (P<0.05). Compared with LPS group, the cell viability of high-concentration luteic acid group increased (P<0.05), and the levels of inflammatory factors TNF-α, IL-1β and IL-6 decreased (P<0.05). Therefore, the high-concentration luteic acid group was selected as the optimal concentration for formal experiment. In the formal experiment, the cell viability and anti-apoptotic Bcl-2 protein expression levels in LPS group decreased compared with those in control group (P<0.05), while the expression levels of inflammatory factors TNF-α, IL-1β and IL-6, apoptosis rate and the expression levels of Caspase-3, Wnt and β-catenin proteins increased (P<0.05). Compared with LPS group, the cell viability and anti-apoptotic Bcl-2 protein expression levels in luteic acid group and inhibitor group increased (P<0.05), while the expression levels of inflammatory factors TNF-α, IL-1β, IL-6, cell apoptosis rate and the expression levels of Caspase-3, Wnt and β-catenin proteins decreased (P<0.05). Compared with the luteic acid group, the change trend of the above indexes in the luteic acid + inhibitor group was more significant (P<0.05), and the change trend of these indexes in the luteic acid + activator group was significantly reversed (P<0.05). Luteic acid may promote the cellular activity of human hPDLSCs induced by lipopolysaccharide and inhibit their apoptosis and inflammation by inhibiting the signal transduction of Wnt/β-catenin signaling pathway. This study can provide a new theoretical reference for the treatment of periodontitis.
Key words: periodontitis; gambogic acid; lipopolysaccharide; periodontal membrane stem cells; Wnt/β-catenin signaling pathway
（Acta Laser Biology Sinica, 2024, 33(6): 567-576）