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Quick Search     Advanced Search   2023 Vol.  32 No.  3 Published: 2023-06-28 1 CONTENTS 2023 Vol. 32 (3): 1-2 [Abstract] ( 549 ) [HTML 1KB] [PDF 1262KB] ( 2053 ) 193 WANG Changke, MA Qiong, CAI Qingyu, SUN Haiyang, TANG Xiaoan, KANG Hongxiang Application of Deep Learning in OCT Medical Images of Skin Abstract: As an important research direction in the field of machine learning, deep learning technology has been infiltrated into various fields of scientific research due to its powerful feature of learning ability and excellent portability. In particular, the achievements in the frontier fields of deep learning have played a positive role in the recognition, classification, segmentation and quantification of medical images. Optical coherence tomography (OCT) technology has the characteristics of non-invasive, high resolution, non-invasive acquisition of micron-level high resolution 3D sectional image of biological tissue, and has been widely used in ophthalmology, vascular, dermatology and other biomedical fields. This paper first introduces the basic principles of OCT and deep learning and the advantages of their combination in the field of skin, and then elaborates the application of deep learning in the field of skin OCT medical images, including the analysis and classification of skin cancer, beauty, fingertip image segmentation, and the qualitative and quantitative evaluation of damaged skin. Key words: optical coherence tomography; medical imaging; deep learning; skin disease; convolutional neural network （Acta Laser Biology Sinica, 2023, 32(3): 193-199） 2023 Vol. 32 (3): 193-199 [Abstract] ( 709 ) [HTML 1KB] [PDF 3330KB] ( 2981 ) 200 LI Hong, GUO Qingyuan, GUO Layang, FU Xingxing Research Progress and Application of Serum-free Culture of Insect Cells Abstract: With the rapid development of the field of bioengineering, cell engineering has been paid more and more attention. Insect cell culture technology, as a new field of bioengineering, has important value in modern biology, among which the genetic engineering research using insect baculovirus as the carrier and insect cells as the receptor has also made breakthrough progress. Insect cells cultured in vitro can express foreign genes efficiently, and insect baculoviruses are harmless to humans and animals, which makes insect cell culture technology being widely used in various fields such as biology, medicine and agriculture. In this paper, the research progress of insect cell culture is reviewed, including the basic components of insect cell culture medium in vitro, the addition of unclear medium, the bioreactor for the large-scale culture of insect cellss and the vector system of insect baculovirus expression and its application. It provides a theoretical basis for the development of serum-free culture medium for insect cells. Key words: insect cells; baculovirus; serum-free culture; mass cell culture （Acta Laser Biology Sinica, 2023, 32(3): 200-207） 2023 Vol. 32 (3): 200-207 [Abstract] ( 698 ) [HTML 1KB] [PDF 2666KB] ( 2696 ) 208 JIANG Shuai, YOU Changqiao, DING Yunyun, ZHANG Hongming, QIN Hong, GUO Xinhong Genetic Diversity Analysis of SARS-CoV-2 Based on Correlation Tests of Microsatellites Abstract: Microsatellites were crucial in the control of SARS-CoV-2 genomic expression, population genetic evolution and host immunity interaction. The NCBI database and microsatellite analysis systems were utilized in this study to screen and test the correlation between the number of different microsatellites and genetic characteristics in the original strain and variants of SARS-CoV-2, as well as to investigate the microsatellite features affecting the genetic diversity of SARS-CoV-2. By bioinformatics analysis, the whole-genome sequence database of SARS-CoV-2 was constructed, and information on the genetic characteristics, microsatellite numbers and relative position distribution characteristics of microsatellites in the sequence database were collected; correlation analysis between the number of various microsatellites and one-sample Wilcoxon signed-rank non-parametric tests were performed by biostatistical analysis. The findings revealed that the quantity, proportion, and relative distributing features of various microsatellites in the original strain and SARS-CoV-2 variants (except Lambda and Omicron) were comparable. The proportion of microsatellites with 2 nucleotide repeats (77%～78%), 3 motif repeats (22%～23%), and 6 bp full-length (73%) were relatively high, and the microsatellite density of ORF3a (0.48/100 bp), E (0.44/100 bp), and N (0.40/100 bp) coding regions were also high. The total average number of microsatellites was negatively correlated with the base content (r: –0.799 6, P: 0.009 7), whereas no significant correlation with the number of base substitutions. This study enriched the research on the genetic diversity as well as the evolutionary mechanism of SARS-CoV-2 in the field of molecular biology and provided new ideas for the prevention and control of the corona virus disease 2019 pandemic. Key words: SARS-CoV-2; microsatellites; genetic diversity; base content bias; population genetic evolution （Acta Laser Biology Sinica, 2023, 32(3): 208-216） 2023 Vol. 32 (3): 208-216 [Abstract] ( 691 ) [HTML 1KB] [PDF 2731KB] ( 2570 ) 218 YOU Shiqi, CHEN Yu, YANG Xueting, LI Yunxuan, QIAO Qing, LUO Ying, YAO Meiying, GAO Luoqing, LIU Xin, WANG Yuequn, WU Xiushan, LI Yongqing Construction of Zebrafish wbp11 Gene Knockout Line Abstract: As a member of the WW domain binding protein family, WBP11 is an important splicing factor involved in the pre-mRNA splicing process. Zebrafish, one of the notable model organisms, contains orthological homologous gene wbp11 in its genome. In order to study the mechanism of WBP11 in the early development in vertebrates, a knockout strain of wbp11 in zebrafish was constructed using CRISPR/Cas9 technology. Firstly, gene knockout target sites were designed on the second exon of wbp11 gene, and sgRNA was synthesized by transcription in vitro. After microinjection technology, F0 generation chimeric zebrafish with wbp11-knockout was obtained. Then, the F1 generation zebrafish by crossing chimeric zebrafish with wild-type zebrafish were identified by genotype, and heterozygous zebrafish were screened out. The sequencing results for exact bands of heterozygous zebrafish showed that there were two strains with different sequence deletion. Heterozygous zebrafish of the F1 generation of the same strain were self-crossed. The homozygous zebrafish was screened out via genotyping, meaning the wbp11 gene knockout strain was successfully constructed. The zebrafish with 48 hours post fertilization showed pericardial edema, heart linearization and skeletal curvature obversed by microscopic white light imaging. These defections may be resulted from the abnormal splicing mechanism caused by the deletion of wbp11 gene. This study opens the way to explore the mechanism of WBP11 gene in the early development in vertebrates. Key words: zebrafish; wbp11; CRISPR/Cas9; gene knockout; heterozygote （Acta Laser Biology Sinica, 2023, 32(3): 217-225） 2023 Vol. 32 (3): 218-225 [Abstract] ( 682 ) [HTML 1KB] [PDF 4173KB] ( 2354 ) 226 LI Man, WU Xiaojuan, LIU Lamei, ZHOU Yan, SI Jin, HE Xiaoxiao A Study on the Preparation and Antioxidant Activity of Vitamin C/Resveratrol Nanocomposite Liposomes Abstract:  By using phospholipid molecules with hydrophilic and hydrophobic properties as matrix materials, the nanocomposite liposomes (Lip@Res/VC) containing both fat-soluble resveratrol (Res) and water-soluble vitamin C (VC) have been prepared through thin film hydration method and have been further used for cellular antioxidant studies. In the preparation of Lip@Res/VC, the optimal added amount of Res and VC were determined by evaluating the encapsulation rate of Res and VC, hydration particle size, polydispersal index and zeta potential of nanocomposite liposomes. The results showed that the hydration particle size of Lip@Res/VC was (92.29±5.41) nm and the encapsulation efficiency of Res and VC were (85.13±5.37)% and (64.73±5.51)%, when the added amount of Res was 4.0 mg and VC was 3.5 mg, respectively. The DPPH (2, 2-diphenyl-1-picrylhydrazyl) and ABTS ［2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt］ free radical scavenging assay in vitro showed that the antioxidant activity of Lip@Res/VC was significantly higher than that of Lip@Res or Lip@VC. Furthermore, L02 cells were selected as model cells to study the intracellular antioxidant effect of Lip@Res/VC. The results showed that Lip@Res/VC is capable of removing intracellular reactive oxygen species (ROS) and protecting L02 cells from oxidative damage induced by hydrogen peroxide. Key words: nanocomposite liposomes; resveratrol; vitamin C; reactive oxygen species; antioxidant （Acta Laser Biology Sinica, 2023, 32(3): 226-233） 2023 Vol. 32 (3): 226-233 [Abstract] ( 684 ) [HTML 1KB] [PDF 4068KB] ( 2577 ) 234 HE Yuwen, TANG Guanyu, LI Chuchu, LI Can, JI Chunxiao, CHEN Tao Inducing Effects of COS on Expression of Antimicrobial Protein ANG4 in Intestine Abstract: Angiogenin 4 (ANG4),  a novel antimicrobial protein with multiple biological functions, such as antimicrobial, anti-inflammation, angiogenesis and regulating the body’s immune response, plays a key role in innate immunity. The expression of ANG4 in neonatal mice intestine is regulated by some factors including microbial signal and developmental stage.  Being an analogue of surface molecules of bacterial cell wall, chitosan oligosaccharide (COS) was selected as a inducer to explore the effects on ANG4 expression and mophology in post weaning mice intestine. qRT-PCR and Western blot analyses showed that the mRNA  and protein levels of ANG4 increased suggesting that COS can upregulate the expression of ANG4. The results of HE staining of mouse intestinal tract showed that the height of intestinal villi, the depth of crypt and the value of V/C increased significantly. It indicates that COS can improve the integrity of intestinal mucosae in some extent through either inducing the expression of some intestinal antimicrobial proteins, such as ANG4, or its antimicrobial and immunomodulation activities, and provide the theoretical basis for the applications of COS and ANG4 in controlling the intestinal diseases in piglets. Key words: ANG4; COS; AMPs; qRT-PCR; Western blot （Acta Laser Biology Sinica, 2023, 32(3): 234-239） 2023 Vol. 32 (3): 234-239 [Abstract] ( 678 ) [HTML 1KB] [PDF 3974KB] ( 2303 ) 240 LI Yi, ZHANG Heng, ZHAO Weibiao Effect of Quercetin on Chondrosarcoma Cell Migration, Invasion and EMT by Blocking JAK2/STAT3 Signaling Pathway Abstract: To investigate the effects of quercetin on human chondrosarcoma cell migration, invasion and epithelial interstitial transformation (EMT) and the regulatory role of Janus kinase 2/signal transduction and transcription promoter 3 (JAK2/STAT3) signaling pathway, human chondrosarcoma SW-1353 cells were cultured in vitro and treated with quercetin of different concentrations for 24 h. The optimal concentration of quercetin (50.0 μmol/L) was selected by cell counting kit (CCK-8) for subsequent experiments. The cells were divided into control group with no intervention, quercetin group with 50.0 μmol/L quercetin, positive control group with 6.0 μmol/L adriamycin, inhibitor group involving 50.0 μmol/L quercetin +50.0 μmol/L JAK2/STAT3 pathway inhibitor AG490, and activator group involving 50.0 μmol/L quercetin +0.5 μmol/L JAK2/STAT3 pathway activator colivelin. After 24 h of intervention, cell morphology, migration, invasion, and expression levels of JAK2/STAT3 pathway and EMT-related proteins were analyzed by inverted microscope, Transwell chamber and Western blot. The experimental results showed that the optimum concentration of quercetin was determined by CCK-8 method to be 50.0 μmol/L. Compared with the control group, the growth of quercetin group were positive control group were inhibited, the cell migration number, invasion number, expression levels of N-cadherin, Vimentin, fibronectin (FN), p-JAK2 and p-STAT3 in quercetin group and positive control group decreased (P<0.05), E-cadherin expression level increased (P<0.05); compared with quercetin group, the change trend of the above indexes in inhibitor group was more significant (P<0.05), while the change of these indexes in activator group was reversed (P<0.05). Quercetin can inhibit the growth, migration, invasion and EMT progression of human chondrosarcoma SW-1353 cells, possibly by blocking the JAK2/STAT3 signaling pathway. Key words: chondrosarcoma; quercetin; JAK2/STAT3 signaling pathway; migration; invasion （Acta Laser Biology Sinica, 2023, 32(3): 240-246） 2023 Vol. 32 (3): 240-246 [Abstract] ( 620 ) [HTML 1KB] [PDF 4253KB] ( 2242 ) 247 WANG Dao, ZHANG Hongbo, LIU Wenbin, CHEN Jianlin Analysis of Structure Features and Epitopes of Chlamydia trachomatis Protease CPAF Abstract: Chlamydial protease-like activity factor (CPAF) is a Chlamydia trachomatis protease, which activity results in dampened host inflammation signaling, cytoskeletal remodeling, and suppressed neutrophil activation. We used series of bioinformatics to analyze and predict CPAF protein, physicochemical properties and hydrophobicity, signal peptide, subcellular localization, transmembrane protein structure, conserved region and N-glycosylation sites and phosphorylation sites, the secondary and tertiary structure, ligand-binding regions and small molecular compounds, B-/T-cell epitopes and the interacting proteins. CPAF, which consist of 601 amino acids, the relative molecular weight is 67 252.57 Da, the theoretical isoelectric point is 5.68. The CPAF protein contains a signal peptide and has non-transmembrane regions. The conserved domain of CPAF protein is located at 3～586 amino acid sequence and belongs to CPAF superfamily. The CPAF protein contains 1 N-glycosylation site and 53 phosphorylation sites. The secondary structure is mainly composed of irregular coiling and α-helix. The CPAF protein contains 14 ligand-binding sites, 20 B-cell epitopes, 4 conformational epitopes and several potential T-cell epitopes, and 5 potentially effective screening small molecule compounds. Meanwhile, CPAF may interact with 10 other proteins including htrA、pbpB、recC and so on. This study provides a theoretical basis for further researching the role of CPAF in the pathogenesis of Chlamydia trachomatis, which could be used as a target for vaccine developments and drug therapies. Key words: Chlamydia trachomatis; CPAF; bioinformatics; structure; antigenic epitope （Acta Laser Biology Sinica, 2023, 32(3): 247-258） 2023 Vol. 32 (3): 247-258 [Abstract] ( 670 ) [HTML 1KB] [PDF 7789KB] ( 2540 ) 258 XUE Xinzi, ZHAO Yue, ZHANG Dan, XU Danying, WANG Xia Construction of the Prognostic Model for Cuproptosis-related Genes Prediction of the Immunotherapy Response in Cervical Cancer Abstract: To explore the prognostic value of cuproptosis-related genes in cervical cancer, the clinical data of cervical cancer patients were downloaded from the TCGA database and randomly divided into training group and validation group. Cuproptosis-related genes were screened out by univariate Cox, LASSO-Cox and multivariate Cox analysis, and the risk model was constructed. Overall survival (OS) of the two subgroups and the entire cohort was analyzed by Kaplan-Meier curve, and the prognostic value of the model was verified by ROC curve and PCA. Univariate and multivariate analyses were performed to evaluate the independent prognostic value of clinical features and risk scores. The biological functions and pathways between the two subgroups were analyzed by gene ontology (GO) and Kyoto encyclopedia enrichment of genes and genomes (KGEE), and the sensitivities of the two subgroups to drugs were further analyzed. Finally, prognostic models of five cuproptosis-related genes (FDX1, ARF1, APP, HSF1, MT1A) were constructed. From the survival curve of risk score, OS in the low-risk group was much higher than that in the high-risk group, and the prognosis was good (P<0.05). By univariate and multivariate Cox analysis, risk score an independent prognostic factor (P<0.001). The predictive power of the prognostic model was demonstrated by receiver operating characteristic curve (ROC) and principal component analysis (PCA). ROC curve analysis was used to assess the sensitivity and specificity of risk scores and other clinical features, such as age, grade, and stage. And the results showed that the prognostic value of risk scores was superior to other clinical features. The results of enrichment analysis showed that gene function was mainly concentrated in extracellular matrix and extracellular structure. According to TIDE algorithm, immunotherapy efficacy of patients in the low-risk group was superior to that in the high-risk group. In addition, significant differences were found in the sensitivity of 24 drugs between the two subgroups. The study established a prognostic risk model composed of 5 cuproptosis-related genes, and proved that the model can accurately predict the prognosis of patients. And patients with low risk score are more likely to benefit from immunotherapy, which provides theoretical basis for clinical individualized therapy. Key words: cervical cancer; cuprotosis related genes; risk model; immunotherapy; prognosis （Acta Laser Biology Sinica, 2023, 32(3): 259-271） 2023 Vol. 32 (3): 258-274 [Abstract] ( 720 ) [HTML 1KB] [PDF 21470KB] ( 2127 ) 272 CHENG Shunzhi, FENG Ling, YANG Qi, ZHANG Ziyu, WU Zhengli, ZHANG Yongqin Effects of Extract of Yacon on Nrf2/HO-1/NLRP3 Pathway on Acute Liver Injury in Rats Abstract: Based on Nrf2/HO-1/NLRP3 pathway, the protective effects and mechanism of yacon extract on acute liver injury (ALI) induced by carbon tetrachloride (CCl4) in rats were investigated. Forty-eight healthy male SD rats were randomly divided into 6 groups: control group, model group, low, medium and high dose group and silymarin positive group, in which ALI model was established with CCl4 and the rats were sacrificed for sampling, and the serum and liver biochemical indexes of rats were detected. HE staining was used to observe the histopathological changes of liver in each group, and ELISA was used to detect the expression levels of inflammatory cytokines IL-6, TNF-α and IL-1β in serum of rats. The relative expression of Nrf2/HO-1/NLRP3 pathway related proteins in rat liver was determined by Western blot. The results showed that compared with normal control group, the expression levels of AST, ALT, ALP, γ-GT, MDA, IL-6 and TNF-α in serum of rats in model group significantly increased (P<0.05 or P<0.01), while the expression levels of SOD, CAT, and GSH in liver tissue were significantly decreased (P<0.05). The pathological injury of rat liver tissue cells was obvious (P<0.05 or P<0.01). Compared with model group, the extract of yacon could decrease the expression levels of AST, ALT, ALP, γ-GT, IL-6 and TNF-α in serum in a dose-dependent manner (P<0.05 or P<0.01), which increase the expression levels of SOD, CAT and GSH and decrease the expression levels of MDA in liver tissue (P<0.05). The degree of liver tissue lesions in rats was significantly improved, which the degree of liver cell necrosis and swelling was significantly reduced, and the inflammatory cell infiltration was significantly improved. Silymarin positive group significantly improved rat liver injury (P<0.05 or P<0.01). Compared with model group, the protein expression levels of Nrf2, GCLC, NQO1 and HO-1 in the extract group and positive control group dose-dependenty increased (P<0.05 or P<0.01). The expression levels of NLRP3 inflammator-related proteins (NLRP3, Caspase1, GSDMD and IL-18) were reduced in a dose-dependent manner (P<0.05 or P<0.01). Yacon extracts have a protective effect on acute liver injury in rats caused by CCl4, in which the effect may be related to regulating Nrf2/HO-1 pathway to improve oxidative stress, reducing lipid peroxidation, removing free radicals in mice, inhibiting inflammatory corpuscle NLRP3 function, reducing the synthesis and release of inflammatory factors and reducing inflammation. Key words: yacon extract; acute liver injury; Nrf2/HO-1 pathway; NLRP3 inflammasome; oxidative stress （Acta Laser Biology Sinica, 2023, 32(3): 272-281） 2023 Vol. 32 (3): 272-281 [Abstract] ( 641 ) [HTML 1KB] [PDF 3644KB] ( 2327 ) 282 NIE Gang, QI Junhua The Expression of GSDMB and GSDMD in Skin Cutaneous Melanoma and Its Correlation with Prognosis and Immune Infiltration Abstract: To investigate the expression of GSDMB and GSDMD in skin cutaneous melanoma (SKCM) and their correlation with prognosis and immune infiltration based on bioinformatics. The expression and variation of GSDMB and GSDMD in SKCM were analyzed by GEPIA2 database and cBioportal database. The prognostic value of GSDMB and GSDMD on SKCM was analyzed by GEPIA2 database. The GSCA database was used to analyze the correlation between the expression level of GSDMB and GSDMD in SKCM and the activity of tumor-related pathways. The TIMER database was used to analyze the relationship between the expression level of GSDMB and GSDMD in SKCM and the infiltration level of immune cells. The results showed that the expression level of GSDMB in SKCM tissue was significantly lower than that in normal tissue, while the expression level of GSDMD was significantly higher than that in normal tissue. In 363 SKCM samples, the total variation rate of GSDMB and GSDMD was 9.64%, with mutations predominating (4.96%). The high expression level of GSDMB and GSDMD was significantly associated with the higher overall survival rate of SKCM patients. GSDMB may activate apoptosis pathway in SKCM, while GSDMD may activate Hormone ER pathway and apoptosis pathway in SKCM. The expression level of GSDMB and GSDMD was positively correlated with the infiltration level of various immune cells. This study shows that the high expression of GSDMB and GSDMD in SKCM patients is related to the higher infiltration level of immune cells and overall survival rate. GSDMB and GSDMD are expected to become biomarkers related to immune cell infiltration and prognosis of SKCM. Key words: skin cutaneous melanoma; gasdermin; prognosis; immune infiltration; bioinformatics （Acta Laser Biology Sinica, 2023, 32(3): 282-288） 2023 Vol. 32 (3): 282-288 [Abstract] ( 700 ) [HTML 1KB] [PDF 3811KB] ( 2386 )

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