31 August 2024, Volume 28 Issue 4

    

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  Study of the Effects of Isorhamnetin on Proliferation and Glycolysis of Oral Squamous Cell Carcinoma Cells

  LIAO Jinzhuang, XIE Li, ZHANG Junqing, WANG Jidong

  Life Science Research. 2024, 28(4): 283-288. https://doi.org/10.16605/j.cnki.1007-7847.2024.01.0110

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  To investigate the effects of isorhamnetin on human oral squamous cell carcinoma (OSCC) cells, the impact of different doses of isorhamnetin on the proliferation of OSCC cell lines CAL27 and SCC9 was detected using both cell viability and colony formation assays, and the effect of isorhamnetin on lactate production and glucose consumption in OSCC cells was determined by glycolysis assay. The changes in glycoly-sis-related signaling pathways after treatment with different doses of isorhamnetin were examined through immunoblotting method. Finally, the xenograft mouse model was used to validate the in vivo inhibitory effect of isorhamnetin on tumors. The in vitro experiments showed that isorhamnetin dose-dependently inhibited the proliferation and glycolysis of CAL27 and SCC9 cells, and this inhibitory effect was related to the sup-pression of key glycolysis rate-limiting enzyme hexokinase Ⅱ (HK2) rather than phosphofructokinase (PFK) and pyruvate kinase M2 (PKM2). The in vivo experiments demonstrated that isorhamnetin inhibited the growth of OSCC and reduced the expression of Ki67 and HK2 in tumor tissue. Overall, the results suggest that isorhamnetin inhibits the proliferation and glycolysis of OSCC, and the inhibitory effect is associated with the downregulation of HK2 expression. 
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  Effect and Mechanism of Xihuang Pill on Inhibiting the Proliferation and Invasion of Liver Cancer Cells

  WU Di, ZHANG Jiaqi, YANG Yuting, LI Xiang, CHEN Guoqiang

  Life Science Research. 2024, 28(4): 289-295. https://doi.org/10.16605/j.cnki.1007-7847.2023.10.0195

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  In order to investigate the effect of Xihuang Pill on liver cancer cells and Yes-associated protein/transcriptional co-activator with PDZ-binding motif (YAP/TAZ) signaling pathway, the sera containing the pill ingredients were prepared by intragastric administration and arterial blood collection in SD rats. The prolife-ration and clonogenesis of liver cancer cells were detected by sulforhodamine B (SRB) staining. The hepato-ma apoptosis was evaluated using flow cytometry and terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL) method. The invasion and migration abilities of liver cancer cells were tested using the Transwell model, and their in vivo proliferation ability was detected using the xenotransplantation model of mouse tumor cells. The YAP/TAZ signaling pathway related proteins and gene expression were ex-plored by Western-blot and fluorescent quantitative PCR. The in vitro results showed that the sera containing pill ingredients significantly inhibited the proliferation, clonal formation, invasion and migration of liver can-cer cells, significantly promoted the liver cancer apoptosis, and significantly inhibited the expression of YAP/TAZ signaling pathway related proteins and genes in a dose-dependent manner. The in vivo results showed that Xihuang Pill significantly inhibited the proliferation of liver cancer cells in a dose-dependent manner. The study demonstrated that Xihuang Pill can inhibit the in vitro and in vivo proliferation, invasion, and metastasis capabilities of liver cancer cells, and may play a role in the YAP/TAZ signaling pathway.
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  Research Progress on the Role and Molecular Mechanism of CARM1 in Malignant Tumors

  LIU Shuyi, MA Zefang, XIAO Sayu, GAO Lu, CAO Mingqi, CHEN Ziwei, ZHANG Xudong, LI Yu, ZHAO Tingting

  Life Science Research. 2024, 28(4): 296-304. https://doi.org/10.16605/j.cnki.1007-7847.2023.06.0170

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  Coactivator-associated arginine methyltransferase 1 (CARM1) is a member of the protein arginine methyltransferase (PRMT) family, also known as PRMT4. In recent years, CARM1 has been widely studied as an auxiliary transcriptional regulator, and found to play an important role in the occurrence and develop-ment of various diseases such as tumors, inflammatory diseases and diabetes. It participates not only in epi-genetic regulation by regulating post-translational methylation modification, but also in gene expression regu-lation through RNA metabolism and metabolic reprogramming. In-depth studies have revealed the regulatory role and molecular mechanism of CARM1 in malignant tumors and showed that its regulatory effects in dif-ferent tumors are not the same. It is still necessary to explore its role in regulatory networks in malignant tumors. Herein, the role and molecular mechanism of CARM1 in malignant tumors were reviewed, with a hope of providing new ideas for related research.
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  Effect of High Glucose Acclimation on the Ability of Umbilical Cord Mesenchymal Stem Cells to Resist High Glucose 

  LIU Chunxiang, ZHANG Tianqi, ZHANG Yi

  Life Science Research. 2024, 28(4): 305-311. https://doi.org/10.16605/j.cnki.1007-7847.2024.01.0112

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  In order to explore the high glucose environment resistance of human umbilical cord mesenchymal stem cells (hUC-MSCs) following high glucose acclimation, and to provide highly adaptive cells and methods for diabetes treatment, the culture media with glucose concentration gradients were used to acclimate hUC-MSCs to passage 5, and the high glucose acclimated cells (HGACs) were harvested. At the same time, the cul-ture media with 1.0 g/L glucose was used to culture hUC-MSCs to passage 5, and the low glucose cells (LGCs) were harvested. A total of 1.0×106 LGCs were cultured for 48 h in the culture media with 4.5 g/L glucose, and the high glucose cells (HGCs) were harvested. The size and proliferation of LGCs, HGCs and HGACs were measured, and the cell phenotype was detected by flow cytometry. Intracellular superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were determined using xanthine oxidase and thiobarbituric acid colorimetric assays, and the expression levels of anti-aging and antioxidant genes were detected by qRT-PCR and Western-blot. The cell size of HGACs was significantly smaller than that of HGCs (P<0.05), and the proliferation of HGACs was faster. The expression rates of CD73, CD90 and CD105 in HGACs were above 95%, and those of CD34 and CD45 were less than 2%. The SOD activity in HGACs was significantly higher than that in HGCs (P<0.05), and the content of MDA in HGACs was lower (P<0.05). Meanwhile, the expression levels of cellular anti-aging gene p21 and antioxidant genes Nrf2, HO-1 and NQO1 were significantly higher than those of HGCs (P<0.05). The results demonstrated that high glucose acclimation can in-crease the activity of hUC-MSCs and make them resistant to high glucose environment.
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  Research Progress of Adipose-derived Stem Cells in the Treatment of Obesity

  ZHANG Quanying, YANG Yang, WANG Yan, CHEN Lingli, LIU Wanqing, LI Xiangyong

  Life Science Research. 2024, 28(4): 312-319. https://doi.org/10.16605/j.cnki.1007-7847.2023.11.0212

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  With the economic growth and living standard improvement, the prevalence of obesity is increasing significantly year by year. Obesity causes not only an imbalance in the body’s energy metabolism, but also dysfunction of the cells that make up adipose tissue. Adipose-derived stem cells (ADSCs) form the adipose tissue and can alter the remodeling and function of adipose tissue. The cells are currently considered to play an important role in the prevention and treatment of obesity. Herein, the biological characteristics, functions and anti-obesity mechanism of ADSCs were reviewed, so as to provide new ideas for the study of clinical prevention and treatment of obesity. 
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  Research Progress on the Mechanism of Synergistic Metabolism Through Tyrosine-DOPA Pathway Between Rumen Microorganisms and Ruminants

  CHEN Qiaoqing, XIE Haibin, HUANG Weidong, DING Luoyang, WANG Mengzhi

  Life Science Research. 2024, 28(4): 320-326. https://doi.org/10.16605/j.cnki.1007-7847.2023.09.0186

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  Tyrosine, an aromatic amino acid, is mostly involved in the research on melanin, and there are few reports on the interaction of tyrosine with rumen microorganisms and ruminant hosts. Relevant studies suggested that tyrosine deficiency leads to a limited growth of rumen microorganisms, including protozoa and bacteria (such as rumen protein-degrading bacteria), while tyrosine supplementation promotes the growth of rumen microorganisms, which coordinate with the body metabolism, affecting the host behavior and health through the tyrosine-DOPA pathway. This article reviews the mechanisms of tyrosine affecting the rumen microbial metabolism and further affecting the host body, with the aim of providing some ideas for improving ruminant behavior and health by regulating fermentation of rumen microbes through tyrosine supplementa-tion in breeding industry.
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  Sedum mexicanum Britt., a Newly Recorded Species of Sedum L. from China’s Mainland

  YE Kang, GE Binjie, XIAO Zhiyong

  Life Science Research. 2024, 28(4): 327-329. https://doi.org/10.16605/j.cnki.1007-7847.2023.05.0161

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  A new record species of Sedum L. from China’s mainland, Sedum mexicanum Britt., which was found in the Dabie Mountains of Anhui Province, is reported. Its morphological features, color photos, distri-bution and application information are provided.
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  Two New Species of Araneus Clerck, 1757 from China (Araneae: Araneidae)

  QIN Xinyu, MI Xiaoqi, LIU Ping

  Life Science Research. 2024, 28(4): 330-335. https://doi.org/10.16605/j.cnki.1007-7847.2024.04.0142

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   Two new species of the genus Araneus Clerck, 1757 from Gaoligong Mountains, Yunnan Province of China are described and named as A. biloculatus sp. nov. (♂♀) and A. papilioformis sp. nov. (♂♀). De-scriptions, photos of type specimens and copulatory organs are provided. Specimens are deposited in the College of Life Sciences, Hunan Normal University.
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  Construction of a Mitochondrial Oxidative Stress-related Prognostic Risk Model for Hepatocellular Carcinoma Based on Machine Learning Algorithms 

  CHEN Keyu, ZHANG Jian, WU Cichun, JIANG Chuan, PENG Shifang, FU Lei

  Life Science Research. 2024, 28(4): 336-347. https://doi.org/10.16605/j.cnki.1007-7847.2023.12.0217

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  Liver cancer is one of the common malignant tumors, and the prognosis for advanced liver cancer is extremely poor. In view of the important role of mitochondrial oxidative stress in the development of hepa-tocellular carcinoma (HCC), mitochondrial oxidative stress-related genes were selected to construct a prog-nostic risk model for HCC. Firstly, the prognostic key genes were screened by using univariate Cox regres-sion analysis and three machine learning methods, namely support vector machine, random forest analysis, and LASSO regression analysis, and a model was constructed based on multivariate Cox regression. Secondly, the prognostic value of the model was further validated in the database. Thirdly, the possible mechanisms of the prognostic differences between the high- and low-risk groups were explored using gene enrichment analysis, and the immune microenvironment and treatment response between the two groups were compared. Finally, the expression of key genes in liver cancer tissues was verified by reverse transcription quantitative real-time PCR (RT-qPCR). Results showed that a total of 10 genes including PDE2A, TREM2, BMP6, NQO1, CPS1, EPO, MAPT, G6PD, SFN and HMOX1 were chosen out. Compared with the low-risk group, the high-risk group of HCC patients had a worse prognosis (P<0.000 1). Enrichment analysis showed that peroxisome proliferator-activated receptor (PPAR) signaling pathway was significantly different between the high- and low-risk groups. And the tumor immunity analysis showed that the tumor immune infiltration, immune check-point-related genes, and immunotherapy response were also significantly different between the two groups. Validation results using RT-qPCR indicated that, compared with normal liver tissues, expressions of CPS1, PDE2A and BMP6 were lower in HCC tissues (P<0.05), while expressions of G6PD and SFN were higher in HCC tissues (P<0.05). In conclusion, the mitochondrial oxidative stress-related prognostic risk model estab-lished in this study has good predictive efficacy and accuracy, and can be used for the precise treatment of HCC. It would have a high clinical application value.
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  Immunoinformatics Design of Epitope-based Subunit Vaccines Against Neisseria gonorrhoeae

  HUANG Yu, HU Xin, LI Ziyue, ZHANG Lei

  Life Science Research. 2024, 28(4): 348-358. https://doi.org/10.16605/j.cnki.1007-7847.2023.10.0200

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  The dominant epitopes from six conserved outer membrane proteins of Neisseria gonorrhoeae (NG) were screeed, multi-epitope subunit vaccines were designed, and the immune responses against them were simulated. Firstly, computer simulation of six outer membrane proteins was performed through the C-ImmSim online server, and the dominant cytotoxic T lymphocyte (CTL) epitopes, helper T (Th) cell epitopes and B cell epitopes were screened from outer membrane proteins with strong CTL, Th cell and B cell immune responses, respectively. Secondly, after the antigenicity, allergenicity, and toxicity analysis of each candidate epitope, candidate epitopes with different arrangement and combinations were connected with antimicrobial peptide and PADRE sequences to construct multi-epitope subunit vaccines. Thirdly, the designed multi-epitope sub-unit vaccines were simulated by computer to evaluate their abilities to induce humoral and cellular immunity. Finally, the candidate subunit vaccines with good immune effect were screened and further evaluated for antigenicity, allergenicity, physicochemical properties, solubility, and secondary structure to ensure that the candidate subunit vaccines meet the vaccine design standards. Through these processes, eight candidate CTL epitopes, 10 Th cell epitopes, and 14 B cell epitopes were selected out, and a total of 64 multi-epitope sub-unit vaccines were designed. Among them, three subunit vaccines had satisfactory immune simulation results, with strong antigenicity, no allergenicity to human body, and no cytotoxicity, were all stable proteins with good hydropathicity and heat resistance, achieved soluble overexpression in E. coli, and had strong affinity with antibodies, all of which met the standards for vaccine design. The results indicated that the bioinfor-matics prediction based mainly on C-ImmSim had a good application prospect in the reverse design of multi-epitope subunit vaccines and high-throughput preliminary screening of their immune effects. This study pro-vided a new and effective approach for the development of N. gonorrhoeae vaccines.
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  Exploration of Ideological and Political Teaching Reform in Pharmaceutical Program: Taking “Lectures on Frontiers of Pharmaceutical Engineering” as an Example

  WANG Yandong, ZHANG Hongli

  Life Science Research. 2024, 28(4): 359-363. https://doi.org/10.16605/j.cnki.1007-7847.2023.09.0193

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  Lack of curriculum ideology and politics is a problem in science and engineering education in colleges and universities, and it is necessary to explore how to integrate curriculum ideological and political elements into teaching practice. “Lectures on frontiers of pharmaceutical engineering” is a professional course for pharmaceutical majors, which is a seminar set up for senior undergraduate students and plays an impor-tant role in the cultivation of pharmaceutical professionals. Herein, the characteristics of the course content were analyzed, and the suitable ideological and political elements were collected, which would be organically integrated into the normal teaching activities through multi-means, multi-level, cross-field, and multi-angle ways. It is hoped that the ideological and political teaching reform will provide ideas for training high-quality innovative professionals in the pharmaceutical field.
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  Exploration of the Path of Integrating Biosafety into Undergraduate Experimental Course of Medical Microbiology Under the Background of New Medicine: Taking Hunan Normal University as an Example

  ZHONG Zhihong, LIAO Minjing, YE Xiangli, YANG Chang, ZHANG Xia, TIAN Mei, LIU Rushi, GAO Peiqi, ZHENG Jiao

  Life Science Research. 2024, 28(4): 364-369. https://doi.org/10.16605/j.cnki.1007-7847.2023.12.0223

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  In recent years, biosafety, especially concerning pathogenic microorganisms, has been widely con-cerned by the society. As medical works are faced with different levels of biosafety risks, it is necessory for relevant staff to have the corresponding knowledge of biosafety and emergency handling ability. Medical mi-crobiology is a compulsory basic course for students majoring in medical laboratory science. Most of experi-mental content of the course involves use of pathogenic microorganisms that may cause uncontrollable trans-mission, thus the laboratory exercises must be carried out in the biosafety laboratory. Previously, the course used to take symbol knowledge as the teaching approach, which can not help students get safety conscious-ness of experiment and embodied experience. According to the requirements of relevant laws and regulations, a biosafety level 2 (BSL-2) laboratory has now been established for undergraduate curriculum in Hunan Nor-mal University. It has been shown that the laboratory exercises conducted in the BSL-2 lab can improve the biosafety consciousness of students and ensure the biological safety in medical microbiology experimental ope-ration through integrating the biosafety awareness training of students, laboratory biosafety system improve-ment, laboratory biosafety management and biosafety assessment into the daily work of experimental teaching.
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  Application of Hypothesis-verification Mode in Experimental Teaching of Ecological Monitoring 

  GUO Chengyuan, ZHANG Jiudong, MENG Changjun

  Life Science Research. 2024, 28(4): 370-376. https://doi.org/10.16605/j.cnki.1007-7847.2024.03.0129

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  The reformed teaching mode was driven by problem hypotheses with practical verification as a starting point, combined with scientific research hotspots such as biodiversity and plant functional traits. The aim of the teaching was to mobilize the students’ subjective initiative in exploring the ecological environ-ment around them, and enhance their problem-solving ability in limited class hours. The experiments in-cluded plant diversity monitoring, the relationship between leaf mass per unit area (LMA) and leaf relative wa-ter content (RWC) and soil moisture content, changes in leaf stomatal traits and photosynthetic pigments in different habitats. This experimental teaching mode had stronger comprehensiveness and practicality, and could be used to train students in their logical thinking, teamwork ability and scientific research quality, to cultivate their ecological awareness and to integrate their theoretical knowledge with practical skills. The ap-plication of the hypothesis-verification mode significantly improved the effectiveness of ecological monitoring experimental teaching, and 76.06% of the students believed that this mode was more dynamic and signifi-cantly raised their learning effect and ecological awareness.