(1. Department of Respiratory and Critical Care Medicine, the Affiliated Jiangning Hospital of Nanjing Medical University, Nanjing 211100, China; 2. Department of Respiratory and Critical Care Medicine, Shanghai East Clinical Medical College, Nanjing Medical University, Nanjing 211166, China)

Abstract: Liquid-liquid phase separation (LLPS) is a biophysical phenomenon within cells that holds significance in cellular regulation, signal transduction, and various biological processes. Over recent years, it has caught considerable attentions. This paper delves into the fundamental concepts, mechanisms of formation, physiological functions, and disease associations related to LLPS. Furthermore, it projects future research trajectories and potential applications. Investigating LLPS offers fresh insights into cellular regulatory mechanisms and disease progression, simultaneously opening avenues for drug development and biotechnological innovations.

Key words: liquid-liquid phase separation; membrane-less organelles; regulatory mechanisms; physiological function; disease development

（Acta Laser Biology Sinica, 2024, 33(2): 097-107）

(1. College of Life Science and Biopharmaceutical Science, Guangdong Pharmaceutical University, Guangzhou 510006, China; 2. School of Health sciences, Guangzhou Xinhua University, Guangzhou 510520, China)

Abstract: Breast cancer ranks as one of the most prevalent malignancies globally, predominantly affecting the female population. Notably, the expression of class IIa histone deacetylases (HDACs), particularly histone deacetylase 5 (HDAC5), exhibits significant variation between breast cancer patients and healthy individuals. This differential expression positions HDAC5 as a promising biomarker for breast cancer and potentially other cancer types. Furthermore, HDAC5 has emerged as a credible molecular target for anticancer therapeutics. This review aims to concisely summarize the structural characteristics of HDAC5, its contributory role in the pathogenesis of breast cancer, and the therapeutic potential of HDAC5 inhibitors. We will discuss the feasibility of detecting HDACs, designing histonedeacetylase inhibitors (HDACi), and identifying effective drug targets in conjunction with HDACi. Our goal is to offer strategic insights for advancing breast cancer treatment, focusing on the application of HDACi in managing breast cancer tumors.

Key words: breast cancer; histone deacetylases; histone deacetylases 5; histone deacetylases 5 inhibitor; drug action targets

（Acta Laser Biology Sinica, 2024, 33(2): 108-114）

 （1. College of Agriculture, Hunan Agricultural University, Changsha 410128, China; 2. Institute of Cotton Sciences of Hunan Province, Changde 415101, China）   

Abstract: Xiang FZ001 was used as a test variety in this experiment, which was conducted in a plant factory with a cycle of 16 hours of light and 8 hours of darkness to study the effects of varying light intensities on the growth and development of cotton (Gossypium hirsutum L.), three light intensity treatments with parameters of L1: 450 µmol·m-2·s-1, L2: 600 µmol·m-2·s-1, and L3: 750 µmol·m-2·s-1, were established to determine the dry weight of each cotton organ. The height of the cotton plant was measured using a straightedge, the chlorophyll content of cotton leaves was measured using a SPAD-502 chlorophyll meter. Additionally, six chlorophyll fluorescence parameters were measured, including initial fluorescence (F0) and maximum fluorescence (Fm), using a Flour Pen 110. The relative chlorophyll content and plant height of cotton were greatest under the L1 light intensity treatment, the root dry weight of cotton was greatest under the L2 light intensity treatment, and the total weight of cotton stems, leaves, and individual plants under the three light intensity treatments were L3>L2>L1 in descending order, and the F0 and Fm were L1>L2>L3 in descending order. The maximum photochemical quantum yield (Fv/Fm) was L2>L3>L1 in descending order. The photochemical bursting coefficient (qp) and effective quantum yield (φPSII) of cotton under the same light intensity treatment showed the same curves of change. During the pre-growth era, there was no significant difference in the qp, φPSII, and non-photochemical burst (NPQ) of cotton under the three light intensity treatments. However, in the late period, there was a greater difference. For the late growth stage, cotton’s qp and φPSII under the three light intensity treatments were high to low in the order of L3>L2>L1, and high to low in the order of L2>L3>L1. High light intensities increase the dry weight, stem dry weight, and leaf dry weight of cotton plants; low light intensities enhance the height of the cotton plant, but too high or too low a light intensity prevents the growth of cotton roots. Compared with cotton under L1 and L3 light intensity treatments, L2 light intensity was suitable for cotton growth, avoiding both inter-plant competition for light and the phenomenon of photoinhibition, with moderate chlorophyll content, resulting in the highest photochemical efficiency of cotton. This study can provide a guidance for cotton factory production and cotton breeding accelerator development and application.

Key words: plant factory; light flux density; Gossypium hirsutum L.; growth and development; photosynthetic characteristics

（Acta Laser Biology Sinica, 2024, 33(2): 115-122）

(School of Information and Communication Engineering, North University of China, Taiyuan 030051, China)

Abstract: In order to investigate the radial farthest detection distance and the appropriate detector arrangement for diffuse reflectance spectroscopy in detecting effective information from internal tissues, a human subcutaneous tissue model was constructed, and the transmission process of 760 nm near-infrared light in the tissues was simulated by using the Monte Carlo (MC) method. Two combined fat-muscle MC models were established according to the optical properties of different tissues. In this model, placing the detector approximately 3 cm, or closer, from the light source allows for the acquisition of valid information about the interior of the tissue. The distribution, number and residual weights of escaping photons at different distances were analyzed in different cases. The results showed that under this model, placing the detector at a distance of about 3 cm, or closer, from the light source can effectively acquire photons inside the tissue. Finally, based on the results of the MC simulation, three detector arrangements that can be applied to noninvasive blood flow or blood oxygenation detection were analyzed and designed. Through comparison, it is found that the hybrid multi-light source-multi-detector arrangement design can not only collect the diffuse light from the tissue more effectively and provide a more uniform photon distribution, but also the design can maximize the number of detectors and realize the detection of a larger area of the same depth of the tissue, compared with the single light source-multi-detector, which provides ideas for the design of spectroscopic instruments for noninvasive detection of near-infrared blood flow or blood oxygenation.

Key words: near-infrared diffuse light; Monte Carlo simulation; human tissue model; source-detector arrangement;  non invasive detection spectroscopic instrument

（Acta Laser Biology Sinica, 2024, 33(2): 123-132）

(School of Electronic and Optical Engineering, Nanjing University of Science and Technology, Nanjing 210094, China)

Abstract: In this paper, based on a simplified dermis model of cylinder-birefringence, the variation of polarization properties of human skin with its structural parameters was studied. To the best of our knowledge, for the first time, the birefringence parameters of human skin measured by polarization-sensitive optical coherence tomography system was used to analyze the changes of birefringence, dichroism and depolarization with the change of structural parameters. The results of this study are important for interpreting the results of polarization experiments, diagnosing human skin diseases and monitoring the effects of treatment in vivo.

Key words: human skin model; birefringence; dichroism; imaging system; depolarization

（Acta Laser Biology Sinica, 2024, 33(2): 133-142）

(Laboratory of Animal Nutrition and Human Health, Hunan Normal University, Changsha 410081, China)

Abstract: β-tectorin, encoded by the TECTB gene, plays an important regulatory role in human hearing function, is an important component of the covering membrane in the inner ear organ, and plays a key role in the process of sound transduction into neural signals. The mutation of β-tectorin can lead to the occurrence of non-syndromic deafness in humans. However, the molecular mechanism of hearing impairment remains unclear. To study the role of the tectb gene in zebrafish inner ear development, this study successfully constructed a zebrafish model with tectb gene deletion using CRISPR/Cas9 gene editing technology and conducted a preliminary analysis of the phenotype. First, target sites and genotype identification primers were designed on exon 3 of tectb gene. Guide DNA (sgDNA) was amplified in vitro, next guide RNA (sgRNA) was obtained by in vitro transcription. Then, sgRNA and Cas9 enzymes were co-injected into wild-type zebrafish embryos by microinjection. After genotyping, the mutant chimera F0 generation fish were screened; and after being raised to adult, they were crossed with wild-type zebrafish to obtain the mutant heterozygotes of the F1. After genetic stability identification and sequencing, F1-generation fish with frameshift mutation were raised to adulthood, they were crossed to obtain F2 tectb homozygotes. Finally, through observation and hair cell staining experiments, it was found that the inner ear structure of the homozygotes mutant zebrafish appears normal, with no obvious difference in the shape and size of the otolith, and no obvious defect in the development of hair cells. However, whether there were changes in the development of internal organs and tissues should be further studied. This study provides us with a research model to gain insight into the molecular mechanisms of inner ear development and human diseases. With the help of the tectb gene knockout zebrafish model, we can elucidate the role of this gene in diseases and better understand the pathways involved in inner ear development, which is conducive to further research on the regulatory mechanism of hearing.

Key words: zebrafish; tectb; CRISPR/Cas9; inner ear development; hair cells

（Acta Laser Biology Sinica, 2024, 33(2): 143-150）

(1. College of Life Science and Technology, Central South University of Forestry and Technology, Changsha 410000, China; 

2. College of Biology, Hunan University, Changsha 410000, China)

Abstract: Extracellular vesicles (EVs) were isolated from the culture supernatant of lung adenocarcinoma A549 cells. The size of EVs was characterized as approximately 100 nm via DLS and TEM analysis, with the EV-specific marker TSG101 detected in the samples. The extracted EV DNA (evDNA) showed a predominantly～12 kb size distribution as assessed by agarose gel electrophoresis. Based on sequencing data, specific evDNA was selected for PCR detection. DNase I or PS nuclease treatment of the EVs confirmed that the evDNA existed on the outside of the vesicle in a linear manner, and poly-dA tails were added to evDNA by TdT enzyme treatment, allowing for their capture by oligo-dT beads and presenting further evidence of their linear surface distribution. Additionally, a TetO-DNA/TetR-GFP visualization system was established to confirm the presence of TetO-DNA in the EVs, which could be visualized by TetR-GFP proteins under fluorescent microscopy. Enrichment of EVs carrying the TetO-DNA was confirmed via flow cytometry and PCR experiments using TetR-GFP protein. These findings collectively suggest that evDNA is distributed linearly on the surface of EVs, providing a basis for further research into their biological functions.

Key words: lung adenocarcinoma cells; extracellular vesicles; evDNA; TetO-DNA; TetR-GFP

（Acta Laser Biology Sinica, 2024, 33(2): 151-159）

(1. Department of Laboratory Medicine, the Second Affiliated Hospital of Guilin Medical University, Guilin 541199, China; 2. College of Medical Laboratory Science, Guilin Medical University, Guilin 541199, China; 3. College of Intelligent Medicine and Biotechnology, Guilin Medical University, Guilin 541199, China)

Abstract: Coxsackievirus A10 (CV-A10) is one of the most common pathogens causing hand, foot and mouth disease (HFMD). The 39 candidate epitope peptides covering the amino acid sequence of the VP1 region of CV-A10 were screened by using the overlapping peptide method and verified by indirect enzyme linked immunosorbent assay (ELISA) and microneutralization inhibition assays. It has been found that candidate epitope P6 (amino acid 39~53 in VP1 region of CV-A10) showed high reactivity with CV-A10 virus serum, and still showed neutralizing inhibition effect at 3.91 μg/mL dilution mass concentration, which was a potential neutralizing epitope. P6 antiserum was prepared by immunizing mice with P6 peptide, and microneutralization test was used to verify its protective effect. The results showed a 1:8.97 geometric mean neutralizing titer for P6 antiserum, which identified P6 as the neutralizing epitope of CV-A10. In order to understand the conservativeness of P6 sequence in CV-A10, the amino acid sequence of the P6 was compared with the representative strains of CV-A10 genotype, and the results showed that P6 was highly conserved in CV-A10. It indicated that P6 is a conserved linear neutralization epitope of CV-A10, which has the potential to resist the infection of various genotypes of CV-A10, and can be used as a target for the development of CV-A10 epitope vaccine. This study aimed to screen and identify linearly neutralizing epitopes on the CV-A10 VP1 protein and to lay a foundation for the development of CV-A10 epitope vaccine and HFMD prevention.

Key words: coxsackievirus A10; VP1 protein; linear neutral epitope; vaccine; hand, foot and mouth disease

（Acta Laser Biology Sinica, 2024, 33(2): 160-166）

(Department of Dermatology and Aesthetic Surgery the Fifth People’s Hospital of Huai’an, Huai’an 223300, China)

Abstract: This study focused on the effects of the differential expression of microRNA-126-3p (miR-126) on migration and invasion of human cutaneous melanoma (CM) cell line C8161, and the role of Janus tyrosine protein kinase 2/signaling and transcriptional activator 3 (JAK2/STAT3) pathway and epithelial-mesenchymal transition (EMT) process were investigated. Dysregulation of miR-126 was achieved by cell transfection, and miR-126-deregulated cells were treated with the JAK2/STAT3 pathway inhibitor AG490 or the agonist Coumermycin A1. Thus, C8161 cells were divided into 5 groups: control group (without transfection or drug treatment), negative control (NC) group (with mimics control transfection, but no drug treatment), miR-126 group (with miR-126 mimics transfection, but no drug treatment), miR-126 + pathway inhibitor group (with miR-126 mimics transfection and AG490 treatment), miR-126 + pathway agonist group (with miR-126 mimics transfection and Coumermycin A1 treatment). Real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression level of miR-126. The expression levels of EMT key proteins and JAK2/STAT3 pathway proteins were determined by Western blotting. Cell viability, migration and invasion abilities were determined by cell counting kit-8 (CCK-8), scratch healing assay and Transwell chamber combined with matrix gel, respectively. Compared with the NC group, transfection with miR-126 mimics significantly increased the expression level of miR-126 in the miR-126 group (#P<0.05), while the relative cell vitality, cell migration rate and cell invasion number significantly decreased (#P<0.05), accompanied with higher protein level of E-cadherin (#P<0.05), and lower protein levels of Vimentin, N-cadherin, fibronectin (FN), JAK2, STAT3, p-JAK2, and p-STAT3, and as well as lower ratios of p-JAK2/JAK2 and p-STAT3/STAT3 (#P<0.05). More importantly, compared with NC group and miR-126 group, the  above indicators of JAK2 and STAT3 protein levels were further changed in miR-126 + pathway inhibitor group (#P<0.05 and &P<0.05), whereas all the above indexes were less changed in miR-126 + pathway activator group (#P<0.05 and &P<0.05), and cell invasion number and FN, JAK2 and STAT3 protein levels in miR-126 + pathway activator group were little different from NC group without significance. Overexpression of miR-126 inhibits cell viability, migration and invasion of human CM cells presumably by blocking EMT process and inhibiting the activation of JAK2/STAT3 pathway. These results were helpful to provide a new theoretical basis for the clinical treatment of human CM and a new strategy for its treatment.

Key words: cutaneous melanoma; microRNA-126-3p; Janus tyrosine protein kinase 2/signal transduction and transcription activator 3; migration; invasion

（Acta Laser Biology Sinica, 2024, 33(2): 167-175）

（School of Basic Medical Sciences, Youjiang Medical University for Nationalities, Baise 533000, China）

Abstract: The aim of this paper is to explore the mechanism of zinc-α2-glycoprotein (ZAG) forming a feedback pathway with extracellular signal- regulated kinase 1/2 (ERK1/2) and p38 to regulate epithelial mesenchymal transition (EMT) in rat renal epithelial cells (NRK-52E) induced by high uric acid environment, we divided NRK-52E into normal control group and high uric acid-induced group which was stimulated by 20 mg/dL uric acid for 48 h, and the cells in each group were transfected with overexpression of ZAG and knocked down respectively, to observe the interactions between ZAG expression level and ERK1/2 and p38 signaling pathways in the cells. The results revealed that the mRNA and protein expression of EMT-related molecules were elevated in rat kidney cells under high uric acid environment compared with the normal culture group (P<0.05); up-regulation of ZAG decreased the expression of mitogen-activated protein kinase kinase (MAPKK), ERK1/2, p38, activated transcription factor-2 (ATF2) and protein kinase B (PKB or Akt) mRNA in this pathway in rat kidney epithelial cells under high uric acid environment culture (P<0.01), while the downregulation of ZAG increased (P<0.05). At the protein level, the expression of MAPKK, p38 and Akt decreased in the ZAG up-regulated group compared to the untransfected group (P<0.05, P<0.01, P<0.001); and the expression of ERK1/2, p38 and Akt was elevated in the ZAG down-regulated group (P<0.001, P<0.05, P<0.01). The results suggest that transfection up-regulation of ZAG decreases the expression of laminin and vimentin mRNA, the marker genes related to EMT in NRK-52E; regulation of ZAG expression in NRK-52E can play a positive role in ERK1/2 and p38 signaling pathways, and thus inhibit renal cell EMT. This study provides an experimental basis for clinical treatment of hyperuricemic nephropathy (HN).

Key words: nephrofibrotic: epithelial mesenchymal transformation of urine; uric acid nephropathy; hyperuricemia; zinc-α2-glycoprotein

（Acta Laser Biology Sinica, 2024, 33(2): 176-184）

(Department of Oncology, Handan First Hospital, Handan 056002, China)

Abstract: To investigate the effects of ginkgolide B on the proliferation and apoptosis of esophageal cancer cells and the related mechanisms. Human esophageal cancer OE19 cells were cultured in vitro. They were divided into control group (no intervention), low/medium/high dose test group (adding 6.25, 12.50 and 25.00 μmol/L ginkgolide B, respectively), ginkgolide B group (12.50 μmol/L ginkgolide B), positive drug group (4 mg/L cisplatin) and inhibitor group ［12.50 μmol/L ginkgolide B+10.00 μmol/L janus kinase 2/transcriptional activator 3 (JAK2/STAT3) pathway inhibitor AG490］ and activator group (12.50 μmol/L ginkgolide B+0.50 μmol/L JAK2/STAT3 pathway activator Colivelin), 24 h after intervention, cell count kit 8 (CCK-8), 5-acetyl-2' deoxyuridine (EdU) method, Hoechst 33258 staining, real-time fluorescence quantitative PCR (RT-qPCR) and protein immunoblot (WB) were used to detect cell viability, proliferation rate, apoptosis rate and expression levels of related factors. The results show that: compared with the control group, the cell viability of the medium/high dose test group and positive drug group was decreased (P<0.05), therefore, in this study, 12.50 μmol/L ginkgolide B group with significant difference and lower concentration was selected as ginkgolide B group for follow-up tests. Compared with the control group, the cell proliferation rate, Cyclin D1 mRNA and protein expression levels, p-JAK2 and p-STAT3 protein expression levels of ginkgolide B group and positive drug group were decreased, while the apoptosis rate and Caspase-3 mRNA and protein expression levels were increased (P<0.05). Compared with the ginkgolide B group, the changes of all indexes in inhibitor group were further enhanced (P<0.05). while those in activator group were significantly reversed (P<0.05). Ginkgolide B can inhibit the proliferation of OE19 cells and promote apoptosis of esophageal cancer cells by down-regulating JAK2/STAT3 signaling pathway. This study revealed a new anticancer mechanism of ginkgolide B and provided a theoretical basis for the study of esophageal cancer.

Key words: esophageal cancer; ginkgolide B; janus kinase 2/transcriptional activator 3; proliferation; apoptosis

（Acta Laser Biology Sinica, 2024, 33(2): 185-192）