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Quick Search     Advanced Search   2022 Vol.  31 No.  2 Published: 2022-05-19 1 CONTENTS 2022 Vol. 31 (2): 1-2 [Abstract] ( 1016 ) [HTML 1KB] [PDF 991KB] ( 3981 ) 97 LIU Huiyu, LI Wen, LIU Zixuan, LI Xiao, HUANG Yanqin Discussion on the Pathogenesis of Type 2 Diabetes: Mitochondria-associated ER Membranes Regulates Endoplasmic Reticulum Stress in Islet β Cell Abstract: Type 2 diabetes mellitus (T2DM) is one of the common clinical endocrine disorders, insulin resistance and insulin deficiency are the two most characteristic pathological features of T2DM. The pathogenesis of T2DM are complex. Endoplasmic reticulum stress (ERS), mitochondrial oxidative stress, apoptosis and other mechanisms were closely related to the pathogenesis of T2DM, whereas the specifically pathogenesis is unclear. The endoplasmic reticulum-mitochondrial structure coupling also known as mitochondria-associated ER membranes (MAM), has a direct or indirect regulation on intracellular calcium (Ca2+) distribution, and the distribution of Ca2+ has great significance to the stability of the intracellular environment. Persistent ERS results in structure damage to MAM, reducing the ability of MAM for regulating the endoplasmic reticulum and mitochondrial to release and uptake Ca2+. Excessive accumulation of Ca2+ in mitochondria activates oxidative stress, exacerbates ERS, and then initiates the apoptotic process of islet β cell. ERS in peripheral tissues can lead to insulin resisitance. The above causes eventually lead to the occurrence and development of T2DM. This article describes the regulatory role of the MAM structure in ERS and its association with the pathogenesis of insulin resistance, and provides the basis to explore the clinical treatment and pathogenesis of T2DM. Key words: type 2 diabetes mellitus; endoplasmic reticulum stress; mitochondria-associated ER membranes; oxidative stress; calcium （Acta Laser Biology Sinica, 2022, 31(2): 097-103） 2022 Vol. 31 (2): 97-103 [Abstract] ( 842 ) [HTML 1KB] [PDF 1552KB] ( 3646 ) 104 HU Jiashen, YIN Huijuan, DONG Xiaoxi, FANG Xiang Effect of Optical Parameters on LED Phototherapy to Inhibit the Proliferation of Common Bacteria in Oral Diseases Abstract: Optical parameters such as wavelength, power density and irradiation time have important effects on the bactericidal effect of phototherapy. This study aimed to explore the optimal optical parameters of LED phototherapy to inhibit the proliferation of oral bacteria, and provide a basis for the formulation of optical therapy for clinical oral bacterial infectious diseases. Enterococcus faecalis and Porphyromonas gingivalis were inoculated in 96-well plates and irradiated with the self-made 405 nm LED and 455 nm LED with power densities of 10 mW/cm2 and 40 mW/cm2. Energy densities of the LEDs ranged from 9.6 to 33.6 J/cm2. After 30 min, the bactericidal effect of phototherapy was evaluated by bacterial fluorescent staining or plate counting. The results showed that 405/455 nm LED had significant bactericidal effects on both kinds of bacteria. The maximum bacteriostatic rate (96.05±1.80)% of P. gingivalis was achieved with 405 nm LED with 40 mW/cm2 at light dose of 33.6 J/cm2. The maximum bacteriostatic rate (97.64±2.31)% of E. faecalis was achieved with 405 nm LED with 10 mW/cm2 at light dose of 33.6 J/cm2. It indicated that LED illumination can achieve significant bacteriostatic effects on common oral pathogens with appropriate optical parameters, while 405 nm LED with light dose of 33.6 J/cm2 could achieve the best effect in the study. Key words: LED phototherapy; endogenous photosensitizer; Enterococcus faecalis; Porphyromonas gingivalis; oral infection （Acta Laser Biology Sinica, 2022, 31(2): 104-113） 2022 Vol. 31 (2): 104-113 [Abstract] ( 839 ) [HTML 1KB] [PDF 7087KB] ( 2659 ) 114 YI Bin, WU Hao, ZHANG Hanxu, GUO Qingxia, HE Jun, SONG Nan, ZHANG Fengmin, SONG Wuqi Mechanism of Red/Near Infrared LED Inhibiting Pain Abstract: To study the inhibitory effect of red/near-infrared light irradiation therapy (LED phototherapy) on pain, the expression of pain related inflammatory factors and chemokines were analyzed in peripheral blood monocytes. Base on evaluating the safe dose of red light/infrared light with human monocyte THP-1, PBMC from peripheral blood monocytes of healthy adults were prepared. After red light/infrared light irradiation, the expression of pain related inflammatory factors and chemokines were detected by fluorescence quantitative PCR. After treatment with red/infrared light, the gene expression levels of CXCL12, CXCL13, CX3Cl1, CCl4, CCR2, IL-4, IL-6 and MMP2 in PBMC in the LED group were significantly down regulated compared with the control group. Red/near infrared light phototherapy can play an antipain role by inhibiting the expression of pain related chemokines and inflammatory factors in PBMC. This study may provide new ideas for pain adjuvant treatment. Key words: red/near infrared light irradiation therapy; peripheral blood monocytes; chemokines; inflammatory factors; pain （Acta Laser Biology Sinica, 2022, 31(2): 114-120） 2022 Vol. 31 (2): 114-120 [Abstract] ( 960 ) [HTML 1KB] [PDF 2361KB] ( 3179 ) 121 HUANG Wenqiang, BU Huitong, PEI Chaozhu, HUANG Mingmin, TAN Yongjun The Anti-tumor Activities of Tumor Antigen Peptide M1-10 Abstract: Based on bioinformatics analysis, a transcription factor FOXM1-derived tumor antigen peptide M1-10 was screened out. Through the experiments of lymphocyte proliferation, interferon gamma release, and cell killing, we found that M1-10 peptide could stimulate the immune memory in vivo and produce better effects when combined with HJURP, MELK, VEGFR1, and VEGFR2-derived four antigens. Tumor pre-immunization experiments were carried out with the mouse models of grafted breast cancer and MMTV-PyMT primary breast cancer. Compared with the control group, M1-10 alone produced significant anti-tumor effects, and M1-10 plus four antigen generated stronger anti-tumor effects. In conclusion, the study developed the tumor antigen peptide M1-10 with a strong immunogenicity, which could result in effective tumor immunity when used alone or combined with other tumor antigen peptides. This conclusion provides further insights into the antitumor mechanism of FOXM1. Key words: tumor antigen peptide M1-10; transcription factor FOXM1; immune memory; tumor immunity; mouse （Acta Laser Biology Sinica, 2022, 31(2): 121-128） 2022 Vol. 31 (2): 121-128 [Abstract] ( 814 ) [HTML 1KB] [PDF 4283KB] ( 2998 ) 129 FU Weixiang, SONG Xiaochao, ZHAO Liying, LU Yiguang, WANG Jundong, ZHANG Jianhai Protective Effect of Riboflavin on Arsenic-induced Toxic Injury to the Heart, Lung and Brain in Mice Abstract: In this experiment, 48 5-week-old mice were exposed to arsenic and riboflavin through drinking water. The control group, the arsenic treatment group (20 mg/L As2O3), the riboflavin treatment group (20 mg/L VB2), the arsenic and riboflavin combined group (20 mg/L As2O3+20 mg/L VB2) were set up to explore the alleviating effect of riboflavin on tissue injury caused by arsenic. The results showed that arsenic treatment caused the heart muscle fiber septa widening, fiber bundles bending, myocardial nucleus shedding, alveolar wall thickening and alveolar cavity shrinking compared with the control group. The number of cerebral cortex cells was significantly reduced and disordered, and the nuclei were dissolved and broken. The addition of riboflavin significantly reduced the damage of arsenic to heart, lung and brain. Therefore, this study confirmed that riboflavin intervention has a significant alleviating effect on the structural damage of heart, lung and brain caused by arsenic, it provides new ideas for the toxicological mechanism and prevention of arsenic poisoning. Key words: riboflavin; arsenic; tissues and organs; toxic injury; protective effect （Acta Laser Biology Sinica, 2022, 31(2): 129-133） 2022 Vol. 31 (2): 129-133 [Abstract] ( 860 ) [HTML 1KB] [PDF 5656KB] ( 2684 ) 134 LIAO Jingrong, MAO Feifei, WANG Xinyun, WANG Ziqi, XU Yuanyuan, XUE Shuangshuang, SUN Yujuan, WANG Yawei Dual-wavelength Decoupling Method of Cell Refractive Index and Thickness Based on Phase Diagram Abstract: In the imaging of living cells, the detection results of refractive index and thickness distribution have always been important parameters in the analysis of cell variation and morphology. In optical phase imaging, due to the coupling effect of RI and thickness of cells, the RI of cells cannot be directly measured, hence the decoupling method of RI and thickness of cells has always been a hot topic in this field. This article focuses on the subject of blood cells based on the free lable three-dimensional phase imaging information to establish dual wavelength phase shift equation, and analyse it by the three-dimensional phase imaging under dual wavelength phase information distribution and gradient distribution characteristics and obtain mathematical result of decoupling of the RI and thickness of homogeneous and nucleated heterogeneous samples by the dual wavelength phase distribution function. The RI and the thickness distribution of the outer cell and the inner core are obtained through the simulation experiments of the single medium model and the spherical outer cell kernel model. The error analysis shows that the method is highly accurate, which verifies the correctness of the method of this article. The proposed method can be used to decouple the RI and thickness distribution of red blood cells and five subgroups of white blood cells, which can provide a new method for the analysis of blood cell abnormality and cell morphology. Key words: phase microscopic imaging; blood cell; refractive index; thickness distribution; decoupling method （Acta Laser Biology Sinica, 2022, 31(2): 134-141） 2022 Vol. 31 (2): 134-141 [Abstract] ( 1056 ) [HTML 1KB] [PDF 6104KB] ( 3286 ) 142 TAN Zhixia, ZHANG Jian, LYU Dan, JIANG Zhigang, YUAN Wuzhou, WU Xiushan, YE Xiangli Construction of the Zebrafish Hand2 Gene-knockout Line by Using CRISPR/Cas9 Abstract: Heart and neural crest derivatives expressed transcription factor 2 (Hand2) is a member of bHLH family, also known as dHand, Hed and Thing2, which can be expressed in human, mouse, chicken, zebrafish, Drosophila and other model animals. Domestic and foreign studies have shown that Hand2 mainly affects the development of the right ventricle and plays a key role in the maintenance of right ventricular progenitor cells and morphogenesis of the right ventricle. In order to further study the mechanism of Hand2 gene involved in heart development, a zebrafish Hand2 gene knockout lines was constructed by CRISPR/Cas9 gene editing technology. According to the analysis of bioinformation website, the optimal two target sites were screened, and primers were designed and synthesized. The cDNA of the Hand2 gene sgRNA was obtained by PCR amplification, and then transcribed into mRNA. The mRNA of the two target sites sgRNA and Cas9 protein were co-injected into the Ⅰ-cell stage of zebrafish embryos. The effectiveness of gene knockout was tested in zebrafish embryos and adults, and it was found that there were base deletions and insertions near the target site, indicating that CRISPR/Cas9 was effective for zebrafish Hand2 knockout. In this study, a knockout model of Hand2 gene in zebrafish was constructed using CRISPR/Cas9 gene editing technology, which laid a foundation for the later study of the mechanism of Hand2 gene in human diseases and the clinical development of novel therapeutic targets. Key words: zebrafish; Hand2 gene; CRISPR/Cas9; gene-knockout; therapeutic targets （Acta Laser Biology Sinica, 2022, 31(2): 142-148） 2022 Vol. 31 (2): 142-148 [Abstract] ( 894 ) [HTML 1KB] [PDF 4512KB] ( 3110 ) 149 JIANG Tingting, WANG Dan, LI Hepei, LI Yanchu Application of Metafer 5 Slide Automatic Scanning System in Auxiliary Diagnosis of Circulating Tumor Cells Abstract: The newly established accurate analysis module for detecting circulating tumor cells (CTCs) (Metafer 5) was used in this study. In order to achieve accurate analysis by Metafer 5, the digital module was optimized. Thirty-three malignant tumors patients were enrolled. Meanwhile, in order to acquire the best application parameters by Metafer 5, we screened the best fluorescent dyes by comparing the characteristics of 4', 6-diamidino-2-phenylindole (DAPI), Rhodamine, fluorescein isothiocyanate (FITC) and TexRed, assessed accuracy and efficiency of CTCs scanning among 10×PA, 10×PN, and 20×PA objective lens and optimized the exposure time of each fluorescent channel. Additionally, CTCs identification was evaluated by both manual examination and Metafer 5 system. According to the data, the results showed that scanning time of CTCs by two objective lens was (114.70±22.08) and (116.50±47.38) min, respectively, and the 10×PA objective lens presented higher accuracy and efficiency. When the exposure time of DAPI, EpCAM, CD45 and CEP8 was adjusted to 0.003, 0.800, 0.040 and 0.068 s, respectively, the clearest and best quality fluorescent images were presented. At the same time, comparing to the manual work, based on the optimal parameters of Metafer 5, not only the time cost was 73.40 min less (P<0.05), but also the accuracy reached 100.00%, which was 8.83% higher than manual work. Above all, the optimized Metafer 5 system could detect CTCs automatically, accurately and quickly in blood sample, which could be used for auxiliary interpretation of CTCs, and could have a future application in clinical practice. Key words: Metafer 5 scan system; CTCs analysis; image processing; artificial intelligence; auxiliary diagnosis （Acta Laser Biology Sinica, 2022, 31(2): 149-156） 2022 Vol. 31 (2): 149-156 [Abstract] ( 881 ) [HTML 1KB] [PDF 1638KB] ( 3169 ) 157 SHI Yao, YIN Meiqiang, WEN Yinyuan, LI Lulu, SUN Min, GAO Zhiqiang Bioinformatics of SiSULTR2.1 Gene in Millet and Its Response to Selenium and Sulfur Abstract: Selenium plays a very important physiological and biochemical role in plants, selenates are actively transported by SULTR carrier proteins. SiSULTR2.1 gene sequence was identified from the millet genome database, bioinformatics analysis of SiSULTR2.1 was performed and expression response of leaves sprayed with Na2SeO4 and Na2SO4 was analyzed. The objective of this study was to investigate the physicochemical properties of SiSULTR2.1 gene and its expression response to Na2SeO4 and Na2SO4 treatment in millet, and to provide theoretical basis for the study of selenium and sulfur transport mechanism of SiSULTR2.1 gene in millet. SiSULTR2.1 was hydrophobic, non-secretory protein, without signal peptide, 70 374.55 Da molecular weight, 656 amino acids and its secondary structure is composed of α-helix, β-folding and random crimp, contains STAS domain and sulfate-transp domain that are characteristic of sulfur transporter. Quantitative real-time PCR (qRT-PCR) analysis showed that SiSULTR2.1 has obvious tissue expression specificity, with the higher expression level in the stems and leaves. SiSULTR2.1 could be detected in all tested tissues of millet plant and expression was induced by Na2SeO4 and Na2SO4 for 0 ～ 96 h, SiSULTR2.1 has a faster response to Na2SO4, reaching its peak at 12 h, and a slower response to Na2SeO4. The results would provide a basis for further elucidating the mechanism of SiSULTR2.1 gene in Se and S transport. Key words: millet; sulfate transporter; expression analysis; Na2SeO4; Na2SO4 （Acta Laser Biology Sinica, 2022, 31(2): 157-164） 2022 Vol. 31 (2): 157-164 [Abstract] ( 855 ) [HTML 1KB] [PDF 2671KB] ( 3539 ) 165 LUO Fei, YUAN Haochen, DIAO Juanjuan Bioinformatics Analysis of Key Genes in Henoch-Schönlein Purpura and Prediction of Traditional Chinese Medicine Abstract: To explore the genes, biological processes and metabolic pathways of miRNA in HSP by analyzing the differential expression of miRNA in bioinformatics Henoch-Schönlein purpura (HSP). The original data set GSE80401 was analyzed by GEO database screening, and miRNA differentially expressed was screened out. miRNA target genes were predicted by miRwalk, TargetScan, miRDB database. Gene ontology and pathway enrichment analysis were performed based on Metascape database, and Cytoscape was used to screen key targets. Finally, the key targets were mapped with Coremine Medical database to screen the potential Chinese herbal medicines for HSP. Based on the GSE sample data, 83 target genes were predicted. The key gene was hsa-miR-3945, enrichment analysis revealed biological processes such as response to lipopolysaccharide, response to inorganic substances, apoptotic signaling pathways, and response to extracellular stimuli, KEGG enrichment showed IL-17 signal pathway, TNF signal pathway, Th17 cell differentiation pathway etc. It is predicted that Guizhi, Fuling, Chishao, Taoren and Mudanpi can be used as potential drugs to treat HSP. The analysis of significant differentially expressed genes and potential core targets promotes the further understanding and exploration of the pathogenesis of HSP, and provides a new direction and clinical basis for Traditional Chinese Medicine treatment of HSP. Key words: Henoch-Schönlein purpura; bioinformatics; differential miRNA gene; hsa-miR-3945; traditional Chinese medicine forecast （Acta Laser Biology Sinica, 2022, 31(2): 165-172） 2022 Vol. 31 (2): 165-172 [Abstract] ( 890 ) [HTML 1KB] [PDF 4620KB] ( 3667 ) 173 JIA Rujiang, ZHAO Shushan, YIN Qingchen, LIU Xiuli, YANG Longlong, WEN Guihai Expression and Prognostic Value of MCM2, MCM4, MCM10 in Pancreatic Cancer and Its Action Mechanism Abstract: The expressions of MCM2, MCM4 and MCM10 in pancreatic cancer and their relationship with prognosis were analyzed based on bioinformatics. The expressions of MCM2, MCM4 and MCM10 in pancreatic cancer tissues were significantly higher than those in normal pancreatic tissues (P<0.05). Expression levels of MCM2, MCM4 and MCM10 mRNA were significantly associated with disease-free survival (DFS) and overall survival (OS) in pancreatic cancer patients (P<0.05). There was significant correlation between DFS and OS in cancer patients (P<0.05). Patients with high expression of MCM2, MCM4 and MCM10 had poorer overall survival (P<0.05). We further studied the mechanism and found that the expressions of MCM2, MCM4, and MCM10 were significantly correlated with certain proteins, and the interacting proteins were MCM5, MCM6, RPA3, RPA1, POLA2, ORC2, ORC3, DBF4, CDC6, GINS2, etc. MCM2, MCM4 and MCM10 are mainly enriched in signaling pathways such as preactivation replication complex, DNA replication, DNA synthesis, DNA replication initiation, nuclear DNA replication, CDC6-associated replication start recognition complex, skin flap transfer, mediated pathways, and body temperature auto-regulation pathways. The high expression of MCM2, MCM4 and MCM10 in pancreatic cancer tissues were significantly correlated with the prognosis of patients. The expression of MCM2, MCM4 and MCM10 have obvious correlation with certain factors, which constitute a functional module, participating in the occurrence and development of pancreatic cancer cells through preactivation replication complex, DNA replication, DNA synthesis, DNA replication initiation, nuclear DNA replication, CDC6-associated replication start recognition complex. The discovery of functional modules of MCM2, MCM4 and MCM10 in pancreatic cancer is expected to provide a new prospect for the targeted therapy of pancreatic cancer. Key words: pancreatic cancer; MCM2; MCM4; MCM10; prognostic analysis （Acta Laser Biology Sinica, 2022, 31(2): 173-179） 2022 Vol. 31 (2): 173-179 [Abstract] ( 797 ) [HTML 1KB] [PDF 4613KB] ( 2910 ) 180 WANG Chao, SHI Huaping, DONG Shuyan, XING Zhi, ZHOU Yali, WANG Jiping, LI Runzhi Cloning and Expression Analysis of caleosin Gene from Perilla frutescens Abstract: Caleosin is a kind of protein involved in oil body synthesis. In this study, the PfClo1 and PfClo2 gene fragments were obtained from perilla transcriptome database. The PfClo2 gene was almost not expressed in various perilla tissues, hence the PfClo1 gene was cloned and analyzed by qRT-PCR to investigate the expression characteristics of the gene PfClo1 in different perilla varieties and its response to drought stress. The results of PfClo1 gene sequence analysis showed that the full-length ORF sequence of PfClo1 gene was 609 bp, encoding 202 amino acids, the theoretical isoelectric point was 5.19 and the hydrophilicity coefficient was 0.678. The PfClo1 protein was located in the cytoplasm from our prediction by using PSORT software. The PfClo1 protein had high similarity with caleosin sequence of Salvia miltiorrhiza and Sesamum indicum by blast homologous sequence alignment analysis, and belonged to a typical caleosin family member. The results of qRT-PCR analysis showed that PfClo1 gene was expressed in flowers and seeds of ‘Jinzisu 1’, and the expression level reached the highest in seeds 30 days after flowering. By analyzing the differential expression of different perilla varieties, it was found that the PfClo1 gene expression was positively correlated with the oil formation of perilla seeds. After the ‘Jinzisu 1’ seedlings were treated with 20% PEG, it was found that the expression level of PfClo1 gene reached the highest at 12 h of stress treatment, which was 6.78 times that of the control. It is speculated that this gene may play a regulatory role in the induction of drought stress in perilla. The results provide a theoretical basis for further exploring the function of PfClo1 gene during perilla seed development and response to drought stress. Key words: Perilla frutescens (perilla); caleosin; gene cloning; expression analysis; drought stress （Acta Laser Biology Sinica, 2022, 31(2): 180-187） 2022 Vol. 31 (2): 180-187 [Abstract] ( 793 ) [HTML 1KB] [PDF 7035KB] ( 2846 ) 188 YANG Boyu, CAI Yi, SUN Luning, LUAN Zedong, PANG Wenyan, WU Xiushan, FAN Xiongwei, JIANG Zhigang Extraction Method and Quality Identification of Genomic DNA from Human Tissue Samples Soaked in Formaldehyde Solution Abstract: Human tissue samples soaked in formaldehyde solution are rare resources in many cases, and the extraction of their DNA is a challenging issue. In this paper, genomic DNA was extracted from four formalin soaked human embryo skin and muscle samples by using an improved method for extracting genomic DNA from samples soaked in formaldehyde solution and a generic genomic DNA extraction kit. The purity and concentration of extracted DNA were determined by UV spectrophotometry, and the DNA solution was subjected to agarose gel electrophoresis and imaging. The results showed that the concentration and purity of genomic DNA obtained by the improved extraction method were better than those obtained by the general method. PCR amplification was carried out with the extracted genomic DNA template. Electrophoresis results showed that the improved extraction method had better PCR amplification effect. Our study provides methodological guidance and a reference for the direction of improvement for the extraction of genomic DNA from precious samples soaked in formaldehyde solution. Key words: formaldehyde immersion; human tissue samples; DNA extraction; PCR detection; agarose gel electrophoresis （Acta Laser Biology Sinica, 2022, 31(2): 188-192） 2022 Vol. 31 (2): 188-192 [Abstract] ( 1038 ) [HTML 1KB] [PDF 1521KB] ( 4087 )

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