Abstract: Herein, plant cell wall structures and their concepts, such as plasmodesmata, primary pit-fields and pits, are discussed, and the correlation between secondary walls and apoptosis of cells is analyzed. In addition, two new terms, “wall holes” and “functional pits”, are suggested, and some confusion about hy-drophilicity of lignin and wall structure of transfer cells is clarified. With regard to plastids, it is pointed out that teaching materials usually overemphasized chloroplasts and neglect other plastids. Particularly, some textbooks misinterpret the yellowing of autumn leaves as the transformation of chloroplasts into chromoplasts, when in fact it is due to the consenescence of chloroplasts.

MicroRNAs (miRNAs) are a class of small RNA molecules that are involved in numerous cellular processes and play a key role in the growth and development of living organisms. Given the importance and structural specificity of miRNAs, they have profound implications for the prediction and assessment of di-seases. In the rapid development of miRNA detection methods, catalytic hairpin assembly (CHA) is a novel thermostatic nucleic acid amplification technology, with high sensitivity, high specificity, simple and conve-nient operation, and no enzyme catalysis in the reaction process. Therefore, it has great potential in miRNA detection. Herein, the principle of CHA technology, and the CHA-based miRNA detection strategy in target identification, signal amplification and signal output were introduced, and the current challenges and pros-pects of this technology were also discussed, aiming to provide some ideas and reference for related research.

A drug target is a specific structure in a cell, tissue or organ that interacts with a drug to make it work. Comprehensive identification of drug targets is essential to understanding the mechanism of action of drugs and their potential side effects. Currently, widely used drug target identification and validation tech-niques, such as activity-based protein profiling (ABPP), compound-centric chemical proteomics (CCCP), re-quire the modification of small-molecule compounds, which may reduce or alter the activity of drug mole-cules. Therefore, advanced techniques that do not require chemical modification of drug molecules, such as drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and thermal proteome profiling (TPP), are becoming important tools for drug target research. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a powerful tool for identification of drug target proteins. This paper reviewed drug target research techniques based on LC-MS/MS and future development of drug target identification.

Neuropathic pain (NP), as a common and refractory chronic pain, is caused by damage or diseases of the somatosensory system. The clinical manifestations are mainly spontaneous pain and induced pain, which bring great distress to patients. The mechanisms involved in NP have not been fully elucidated, and it has been widely accepted that peripheral and central sensitization after a nerve injury play critical roles in induction and maintenance of NP. Peripheral sensitization is mainly related to ectopic discharge induced by sympathetic sprouting associated with changes in related ion channels, peripheral inflammatory responses, transcriptional disorders of genes related to non-coding RNAs, and changes in pain signal transduction, while central sensitization is closely related to inflammatory responses, abnormal activation of glial cells and dysfunction of central nervous system. Herein, the mechanisms underlying peripheral and central sensitiza-tion resulted from nerve injuries were reviewed, hoping to provide new ideas for basic research on NP and its treatment.

The hair color comes from melanin. Hair pigmentation can be divided into 3 relatively indepen-dent stages: proliferation and differentiation of melanocytes, synthesis of melanin in melanocytes, and mela-nin transportation. The regulation mechanisms of these 3 stages are different. At present, the substances that promote the generation of hair melanin mainly include hormones, nutrients, antioxidants, and other drugs with various action mechanisms. This paper reviews the research progress of the mechanism of melanin syn-thesis and drugs for treating premature gray hair, analyzes some unclear and important problems involved in hair color formation, and the prospects of related drugs, in order to provide better understanding of mecha-nism of hair color formation and find more effective measures for preventing premature gray hair.

Environmental stimuli play a key role in the survival and evolution of wild animals. Cell receptors that assist in perception, including touch and temperature, reflect biodiversity. Merkel cells (MCs) are impor-tant sensory cells in touch-sensitive areas of vertebrates. Herein, literature research and data statistics were used to investigate the distribution of MCs with the help of PubMed, ScienceDirect and China National Knowledge Infrastructure (CNKI). The results showed that MCs had been observed in 77 species within 6 ani-mal classes including Mammalia, Amphibia, Aves, Reptilia, Cyclostomata and Actinopteri. The histopatholo-gical studies of MCs mainly focused on Mammalia, which accounted for 50% and mainly included human, monkey, rat, mouse, dog, cat, pig and rabbit, then followed by Amphibia and Actinopteri. In tissues and organs, MCs were found to be distributed in nose/vibrissae in 33 species, which accounted for 40%, suggesting that the nose and vibrissae are important parts for animals, especially wild animals, to perceive external mecha-nical stimuli. MCs were also studied in palate, perioral area, fingers/palms of hands/feet, lip, tongue, dorsal skin, gingiva, limbs, abdomen skin, eyes, cheeks, gills and other touch-sensitive sites of many species. Sys-tematically summarizing the distribution of MCs found in known species and studying the distribution of MCs in different species would be helpful to understand the role of environmental diversity in biodiversity.

Catalase (CAT) is a hydrogen peroxide (H2O2) scavenging enzyme and plays a central role in plant growth, development and stress response. Its function is strictly regulated to keep an appropriate concentra-tion of H2O2 as a signal molecule under normal conditions to ensure plant growth and development, and to maintain H2O2 homeostasis under stress conditions to enhance plant stress tolerance. This review summarized the recent research progress of CAT in plant growth, development and stress response, especially the regula-tion of CAT function by post-translational modification and subcellular localization. It also made suggestions for future research on the regulatory mechanism of CAT in plants.

NKX proteins are the most important type of NKL homeobox transcription factor subclass and have been found to be widely involved in regulating cell fate and body pattern formation during embryonic development, especially embryonic development of nervous system, and involved in the occurrence of thyroid cancer, lung cancer, prostate cancer and other tumors. In view of the important roles of NKX proteins in embryonic development and tumorigenesis, the research progress of the structural properties, physiological mechanisms, and disease correlation of 17 proteins in the 7 NKX homeobox protein families were systemati-cally summarized, which would provide helpful information on gene targeted therapy of related diseases. In addition, the lack of genomics research and animal model construction of diseases related to some NKX genes were also discussed, which should be directions for future research.

The OCA2 gene is an important mammalian pigment gene that is involved in the melanin synthe-sis reaction, and is also a main regulator of human eye color. It encodes a 110 kD-sized transmembrane protein, which is located at the melanosome vesicle membrane. Studies have shown that OCA2 affects the melanin synthesis through regulating maturation and pH value of melanosomes, and participating in the first step of the melanin synthesis pathway. The rs12913832 enhancer of the adjacent gene HERC2 modulates the expression level of OCA2 to ultimately control human eye color. In addition, OCA2 is also the causative gene for oculocutaneous albinism typeⅡ(OCA2) and has indirect effects on diseases including Prader-Willi syndrome (PWS), Angelman syndrome (AS) and melanoma, with which patients usually show hypopigmenta-tion of skin or hair. Herein, the research progress of the OCA2 gene from the perspectives of molecular bio-logy, cell biology and genetics is reviewed, so as to provide novel ideas for further exploring the molecular mechanisms of OCA2 and the diagnosis and treatment of the related diseases.

Abstract: Ferroptosis is an iron-dependent non-apoptotic regulated cell death (RCD) mode, which is charac-terized by the accumulation of intracellular lipid peroxidation products and reactive oxygen species (ROS). In recent years, ferroptosis has attracted much attention in the pathogenic mechanism of postoperative cogni-tive dysfunction (POCD), and is closely related to neuroinflammation, mitochondrial energy metabolism, auto-phagy and other pathogenic mechanisms. Herein, the biological characteristics and functions of ferroptosis and its progress in POCD research are reviewed, hoping to provide the latest valuable information for pre-vention and treatment of the neurological disease.

Golgi protein 73 (GP73) is a glycosylated transmembrane protein located in the cis-Golgi mem-brane, which plays an important role in the development of tumors and is a potential target for tumor therapy. Current studies have shown that GP73 can be used as a serological marker to assist the diagnosis of hepatocellular carcinoma (HCC). With the further study of GP73, microRNAs (miRNAs) related to GP73 have been gradually explored and found to be closely associated with the occurrence and development of various tumors. miR-212 and miR-27a can inhibit the invasion and metastasis of HCC, and GP73, miR-27b and miR-493-5p can be used as biomarkers for prognosis of HCC patients. Therefore, GP73-related miRNAs are promising targets for the diagnosis and treatment of liver cancer. Herein, the roles and mechanisms of GP73 and its related miRNAs in the development of HCC are summarized, hoping to provide ideas for the tumor pathogenesis, diagnosis and treatment.

Cognitive functions include learning, memory, attention, and reaction abilities. Learning and memory abilities are easily damaged under low oxygen conditions. Neurons are basic working units for trans-mission of various signals, and synapses are critical parts in signal transmission. If neuroplasticity is im-paired, the integrity and accuracy of neuronal information transmission will be affected, thereby affecting cognitive function. In various low oxygen environments, the production and accumulation of hypoxia-in-ducible factor 1 (HIF-1) and reactive oxygen species (ROS), and the abnormality of apoptosis-related fac-tors, tau proteins and amyloid β-protein (Aβ), can all cause damage to the cerebral cortex and hippocampus, affecting neural plasticity and leading to learning and memory impairments. Currently, the mechanism of low oxygen-induced learning and memory impairment is not yet clear. Herein, the research progress in the mechanism of hypoxia-induced learning and memory impairments and the impact of low oxygen on neural plasticity are reviewed, so as to reveal the possible influencing factors of hypoxia-induced learning and memory dysfunction and provide new ideas for related research.

A preliminary investigation of the species, distribution and floristic characteristics of Impatiens in Guizhou Province has been carried out for more than ten years through field investigation, relevant literature review and specimen research. The results obtained are as follows: 1) There are 69 species (including one variety) of Impatiens L. in Guizhou. They are obviously distributed in narrow range with endemism, and grow more in the south of Guizhou than the north; 2) In terms of geographical composition, most of the Impatiens plants in Guizhou Province are region-specific and belong to tropical Asia and temperate Asia distribution types. Among them, 62 species are endemic to China (89.86%), 10 endemic to Southwest China (14.49%) and 10 endemic to Guizhou (14.49%); 3) Due to their primitive and evolutionary taxa and the characteristics of transition from tropical to temperate zones, the Impatiens plants in Guizhou are probably parts of the evolu-tionary center of Balsaminaceae in China. This investigation is beneficial to understanding the species, geo-graphical distribution pattern and floristic components and characteristics of Impatiens in Guizhou. It also provides reference for studying plant diversity in this province.

Acinetobacter sp. WML1 was isolated from the fishery sediments of freshwater lakes and proved to have application value due to its ability to degrade chitin. Herein, the biosafety of the chitinase-producing strain WML1 was first evaluated for application. The drug resistance of the strain was detected with the disk diffusion method, haemolytic activity was analyzed by streaking the blood agar plate, amino acid decarboxy-lase and nitrate reductase activities were measured by the chromogenic assay with bromocresol purple and nitrate reductase activity assay kit, respectively. On the basis of biosafety analysis, the fermentation condi-tions of the strain were then optimized by single-factor and orthogonal tests for chitinase production. The re-sults showed that the strain WML1 was sensitive to most antibiotics, and had no haemolytic activity or ni-trate reductase activity, had arginine decarboxylase activity and weak lysine and ornithine decarboxylase ac-tivities. The optimal enzyme production conditions of strain WML1 were as follows: 25 mL medium containing 0.4% soybean powder, 0.20% galactose, 0.03% KH2PO4, 0.07% K2HPO4, 0.002% FeSO4, 0.05% MgSO4·7H2O, 0.001% ZnSO4, 0.60% colloidal chitin in a 250 mL flask, with the initial pH of 9.5, 5.0% inoculum size and culture temperature of 37 ℃. Under the optimized conditions, the chitinase activity reached (2.72±0.09) U/mL. In conclusion, Acinetobacter sp. WML1 is generally safe in application, with only low toxicity that is preven-table and controllable, and the enzyme activity could be about 4.61 times higher than that before optimiza-tion. The study provides a theoretical foundation for the industrial application of chitinase-producing bacteria.

Heat shock protein 20 (Hsp20) family contains a kind of functional proteins that are widely syn-thesized in plants in response to abiotic stresses. Herein, the Hsp20 gene family was analyzed by bioinfor-matics using Arabidopsis thaliana genome database. HMMER search and protein physicochemical property analysis showed that there are at least 30 Hsp20 genes in A. thaliana, encoding 14.6~41.4 kD molecular weight proteins, all of which contain an α-crystallin domain (ACD). Phylogenetic analysis showed that 22 of the 30 A. thaliana Hsp20 members belong to 12 different subfamilies, and the remaining 8 belong to un-known taxa, which may be Hsp20-like proteins. Collinearity analysis showed that fragment replication and tandem replication are the main amplification events of Hsp20 members, and most Hsp20 members contain no or only one intron. Analysis using MEME showed that motifs 1, 2, and 9 are common conserved motifs in Hsp20 family. Transcriptome analysis indicated that the expression levels of 15 Hsp20 genes were up-regu-lated after drought and (or) salt stress, which was verified by qRT-PCR assay. These results provide theoreti-cal basis for further investigation of the biological functions of Hsp20 genes.

In order to identify novel cold-tolerance genes in rice, genome-wide gene expression profiling of Chaling common wild rice (CWR) and the cultivar indica rice Peiai 64S subjected to cold stress was perfor-med at the seedling stage using the GeneChip Rice Genome Array. A gene, designated OrCrGHl (Oryza rufi-pogon cold responsive glucan hydrolase-like), was screened from Chaling CWR as a cold-tolerance candi-date, which was highly responsive to cold stress in Chaling CWR but not in Peiai 64S. The expression pro-file of OrCrGHl obtained by the microarray analysis was confirmed by real-time quantitative PCR, suggesting that OrCrGHl is a cold-responsive gene in Chaling CWR. Its full open reading frame (ORF)-containing cD-NA clone was obtained by reverse transcription PCR (RT-PCR). Its sequence analysis showed that it encodes a glucan hydrolase-like protein of 558 amino acid residues, with a molecular weight of 59.252 kD and pI value of 5.81. The deduced amino acid sequence was completely identical with that of the glucan endo-1,3-beta-glucosidase GII isoform X2 of japonica rice Nipponbare, and was 99.28% identical with that of hypo-thetical protein OsI_26347 of indica rice 93-11. Analysis of the putative promoter region revealed several cis-acing regulatory elements that may be related to stress or stress hormone responses. The results indicated that OrCrGHl is a novel candidate gene associated with cold tolerance in wild rice. Further study is in progress to understand more about the gene.

Helicobacter pylori is a Gram-negative, microaerobic bacterium that colonizes the human gastric mucosa and is significantly associated with the development of chronic gastritis, peptic ulcer, gastric mu-cosa-associated lymphoma, gastric tumors, and gastric polyps. Infection of gastric mucosal cells with H. py-lori can activate nuclear factor-κB signaling pathway, induce interleukin-8 (IL-8) release, recruit antigen-presenting cells (APCs), and stimulate the host to produce innate and adaptive immune responses, resulting in anti-H. pylori effects in the host. However, the bacterium has the ability to manipulate and disrupt the host immune system, not only by modifying its own structure to avoid recognition by the host immune sys-tem, but also by inhibiting the proliferation and maturation of T and B lymphocytes, promoting T cell apop-tosis, affecting the balance of Th17/Treg differentiation, and inducing the formation of immune tolerance. Furthermore, H. pylori can inhibit Th1 cell-mediated immune responses and promote immune escape by taking up cholesterol, disrupting the cholesterol-rich microdomains of gastric mucosal epithelial cells, and blocking the assembly of interferon-γ (IFN-γ), IL-22 and IL-6 receptors. This paper summarized the H. py-lori-related immune response and immune escape in humans, in hopes of providing reference materials for clinical control and immunotherapy of H. pylori infection.

Cell biology is a basic course for biological science majors in normal colleges and plays an impor-tant role in professional guidance. Herein, combined with the blended teaching practice of cell biology based on the Rain Classroom, teaching methods, implementation process, resource construction, assessment evalua-tion and educational effects of this course are expounded and discussed. The results of the questionnaire survey showed that the blended teaching mode based on the Rain Classroom can help students better under-stand theoretical knowledge, improve their comprehensive ability and cultivate their interest in learning cell biology.

Melatonin (MT) is a neurohormone released by the body. It has anti-infection, anti-inflammatory and antioxidant effects, and regulates the body’s normal sleep cycle. As an effective free radical scavenger and immune enhancer, it plays an important role in maintaining the normal physiological functions by acting on all systems and tissues of the body. Pyroptosis is a type of programmed cell death, in which the inflam-masomes and GSDMD (gasdermin D) family are the most important components. When the inflammasomes and the GSDMD protein are activated, membrane pores in the cell surface are generated, and a large amount of cellular fluid and intracellular substances are released, leading to a series of inflammatory cascade reactions, followed by secondary cell damage. Pyroptosis is closely related to the occurrence of many diseases. Mela-tonin, as an antioxidant and anti-inflammatory neurohormone, can cope with occurrence and development of myocardial damage, diabetes, central nervous system disease, obesity and degenerative disease by inhibiting pyroptosis. Herein, the effects of melatonin on pyroptosis in different diseases are reviewed.

Birt-Hogg-Dubé (BHD) syndrome is an autosomal dominant genetic disorder characterized by der-matofibroma, renal tumor, pulmonary cyst and spontaneous pneumothorax. At present, diagnosis of BHD syn-drome is based on clinical manifestations, imaging features and genetic evidence. Mutations in Folliculin (FLCN), a tumor suppressor gene, are the main genetic lesion that causes BDH syndrome. The FLCN protein can form a complex with FLCN-interacting protein 1 (FNIP1), FNIP2 and adenosine monophosphate-activated protein kinase (AMPK), acting on occurrence and development of BDH syndrome by regulating mammalian target of rapamycin (mTOR) pathway. Recent studies have found that obvious differences exist in BHD syn-drome-associated renal tumors, sporadic renal tumors and other genetically related renal tumors. Herein, the molecular mechanism and pathological characteristics of BHD syndrome-associated renal tumors were sum-marized, and the differences between BHD syndrome-associated renal tumors and other renal tumors were discussed, hoping to give some reference in clinical diagnosis of BHD syndrome-associated renal tumors and development of effective targeted therapies in addition to surgery.

Non-coding RNAs (ncRNAs), which are abundant in organisms, have been of special concern in recent years. Most ncRNAs do not encode proteins, but they play a significant role in participating complex signaling cascades in various diseases by regulating gene expression and protein functions. Pancreatic cancer is known as the king of cancer because its early symptoms are not obvious, diagnosis often occurs at an ad-vanced stage of the disease and its treatment effect and prognosis are extremely poor. There is still a long way to go in scientific research of pancreatic cancer. This article describes the mechanism of ncRNAs in the occurrence, progression and drug resistance of pancreatic cancer, hoping to provide some ideas for re-searchers to find targets for the tumor detection and treatment.

The spatiotemporal specific expression of genes plays an important regulatory role in organism development and disease occurrence. N6-Methyladenosine (m6A) is the most common epigenetic modification in eukaryotic RNAs. The m6A modification ensures timely and accurate gene expression by changing RNA structure and the interaction between RNAs and RNA-binding proteins, and regulating RNA splicing, sub-cellular localization, translation and stability. Recent studies have shown that m6A not only plays an impor-tant role in the development of organisms, but also is involved in the occurrence of various diseases by changing cell functions. This review focused on the function and mechanism of m6A modification in mam-malian physiological development and disease occurrence, hoping to provide a theoretical basis for using m6A modification in clinical treatment.

Accurate species identification is a prerequisite for biodiversity conservation and management. Mangshan Mountain, located in the south of Hunan and at the border of Hunan and Guangdong provinces, is a priority area for biodiversity protection in Nanling Mountains, with rich diversity of amphibian species. Ac-cording to the Fauna Hunan: Amphibia, there are two Boulenophrys species distributed in Mangshan: B. kua-tunensis and B. brachykolos, but with the further study of the systematic classification of subfamily Mego-phryinae, the distribution of the two species in Mangshan is currently controversial. During May to June, July to August, November 2021 and June to August 2022, four field surveys on the species of Boulenophrys in Mangshan National Nature Reserve, Hunan Province were conducted, and 20 specimens were collected. Through morphological and molecular identification, these specimens proved to belong to four species: B. nanlingensis, B. cf. ombrophila, B. jiulianensis, and B. shimentaina, and the latter three species are new records in Hunan Province. However, the species B. kuatunensis and B. brachykolos were not found to exist in Mangshan. Therefore, the existence of the two species in Mangshan should be further confirmed. In addi-tion, the morphological characteristics of female B. shimentaina and B. nanlingensis were described for the first time, which were lack in the original new record description.

The goal of moving faster to build a powerful agricultural country is proposed in the report of the 20th National Congress of the Communist Party of China. For agricultural colleges and universities, it is im-portant to make sure that the students are well qualified for modern food industry. Food biochemistry is one of the basic professional courses of food science and technology. However, there are problems in the teaching process, such as lack of close connection between courses, insufficient ability for students to transfer know-ledge, disconnection between teaching and engineering education, a low growth value for students and so on. To overcome these weaknesses in teaching and to carry out the engineering education professional certifica-tion, the course content and a new curriculum system were restructured. The teaching reform included inte-grated ideological and political education, innovated teaching models and methods, and diversified assess-ment and evaluation system. These measures proved to bring benefits to students in improving their innova-tion skills, establishing correct scientific emotions and sense of value. In conclusion, the attempts that made in teaching the course have raised the teaching quality, and may provide reference for other course reforms.

Exosomes are nano-sized lipid bilayer vesicles that are fused by the cell membrane and multi-vesicles and released into the extracellular space by exocytosis. Exosomes play an important role in intercel-lular communication. MicroRNAs (miRNAs) are a class of endogenous non-coding RNAs consisting of 18~24 nucleotides. miRNAs play an important role in the regulation of post-transcriptional gene expression. Pul-monary fibrosis is an irreversible and progressive chronic lung disease. Many studies have shown that abnor-mal expression of exosomal miRNAs is closely related to the development of pulmonary fibrosis. This review focuses on the biological characteristics of exosomes and miRNAs, and discusses the important roles of exo-somal miRNAs in pulmonary fibrosis and their potential as a novel biomarker in this disease.

To screen immune infiltration related genes in osteoarthritis (OA), multiple microarray data were obtained from Gene Expression Omnibus (GEO) database, and divided into a training set and a validation set to screen out the differentially expressed genes (DEGs). Then all genes in the training set were analyzed using Gene Set Enrichment Analysis (GSEA) software for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Subsequently, GO and KEGG analyses of DEGs were performed, and a pro-tein-protein interaction (PPI) network of the DEGs was constructed to screen hub genes. The validation set was used for differential validation of hub genes and determination of their diagnostic value. The content and proportion of 28 immune cells in the OA samples were obtained using the single sample gene set enrichment analysis (ssGSEA) algorithm, and the correlation between key gene expression and immune cell infiltration was calculated by Spearman’s correlation analysis. There were 27 DEGs found between the OA and normal car-tilage tissues. The GSEA results showed that gene expression changes in the OA samples were mainly in-volved in the processes such as humoral immune responses and histidine metabolic signaling pathways. GO and KEGG analyses of DEGs showed that the DEGs were associated with signaling pathways related to Staphylococcus aureus infection and NOD-like receptor signaling pathways. Four hub genes CAMP, S100A8, DEFA3, and COL5A2 were screened out from the DEGs, and their abnormal expressions in the OA tissues and high diagnostic value were confirmed. Further analysis of immune cell infiltration showed that obvious changes occurred in activated B cells and other immune cells in the OA cartilage tissues, and the four hub genes were significantly correlated with immune infiltration. The results indicated that activated B cells and other immune cells play an important role in the occurrence and development of OA, and CAMP, S100A8, DEFA3, and COL5A2 are hub genes associated with immune infiltration, which are closely related to the occurrence and development of OA.

In order to explore the hub genes for the difference of adipogenesis and lipid metabolism in dif-ferent tissues of Laiwu pigs, Laiwu pigs with similar body weight at 180 days of age were used to identify the mRNA expression profiles of intramuscular fat (IMF) and subcutaneous fat (SCF). The functions of diffe-rentially expressed mRNAs were analyzed through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). A protein-protein interaction (PPI) network of the top 300 mRNAs with a high average ex-pression level was constructed, key regulatory mRNAs were screened, and sequence features and protein structures of the key mRNAs were analyzed. Meantime, the reliability of the sequencing data was verified by real-time fluorescence quantitative PCR (qRT-PCR) for 6 randomly selected mRNAs. A total of 1 665 differen-tially expressed mRNAs were identified from IMF and SCF mRNA expression profiles, of which 888 were up-regulated and 777 were down-regulated. The GO results showed that the differentially expressed mRNAs were mainly involved in regulating signal pathways in biological process (BP); composing biological membranes in cellular component (CC); affecting the enzyme activity in the cell metabolism process in molecular function (MF). The KEGG enrichment results showed that the differentially expressed mRNAs were mainly involved in adipocyte production, lipid metabolism, inflammation and cancer-related signaling pathways. Through analysis of the PPI network, gene sequence and protein structure characteristics, it was found that C-X-C motif chemokine receptor 4 (CXCR4), platelet and endothelial cell adhesion molecule 1 (PECAM1), urokinase-type plasminogen activator (PLAU), angiotensinogen (AGT), AKT serine/threonine kinase 2 (AKT2), heat shock cognate 70-kD protein, tandem duplicate 2 (HSP70.2), galectin 3 (LGALS3), and periostin (POSTN) were in the center of the regulatory network. The qRT-PCR verification results were consistent with the sequencing results, proving the reliability of sequencing results. By comparing the difference between IMF and SCF tran-scriptomes, it was found that AGT, CXCR4, HSP70.2 and PLAU genes play critical roles in the lipid me-tabolism process of IMF deposition and can be used as candidate genes in IMF regulation.

Programmed death ligand 1 (PD-L1) has emerged as an important target in immune checkpoint blo-ckade therapy for cancer. It is expressed in a variety of cells. Its high expression in tumor cells can enhance programmed death 1 (PD-1) inhibitory signals, thus promoting tumor immune escape. In recent years, cancer immunotherapy based on anti-PD-1/PD-L1 antibodies has brought revolutionary changes in cancer treatment. However, cancer immunotherapy produces long-lasting treatment effects only in some people with cancer. For most patients, it has short-term or no effects. Studies have found that the degradation of PD-L1 is critical for treatment response to cancer immunotherapy. Herein, the lysosomal and proteasomal pathways of PD-L1 degradation and the interaction of PD-L1 degradation with cancer immunotherapy are reviewed, aiming to provide new ideas for improving response rate and scope in cancer immunotherapy.

Clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9 (CRISPR/Cas9) technology is currently widely used in both basic and clinical application research of medicine. Since vectors play an important role in CRISPR/Cas9 technology, further developing and optimizing vector systems is of great significance. Most of the traditional vectors are viral vectors, which have high delivery efficiency, but with defects such as limited insert size, induction of immune response, carcinogenicity, difficulty in large-scale production and even off-target. Non-viral nanocarriers can solve, to some extent, the potential toxicity and capacity limitations caused by viral vectors during gene editing, and may have broader application pros-pects. This paper reviews the non-viral nanocarriers currently used for CRISPR/Cas9 system delivery, inclu-ding main difficulties that non-viral nanocarriers may encounter in delivering CRISPR/Cas9 systems, and cor-responding solutions and strategies, in order to provide a new reference for gene therapy and drug development.

Parkinson’s disease (PD) is a serious threat to the health of middle-aged and elderly people. The etiology of the disease remains unclear. Recent studies have found that the occurrence of PD is closely related to the intestinal microbiome, which is involved in the process of PD by affecting the intestinal barrier permea-bility, inflammation and oxidative stress levels and the release of neurotransmitters through the microbe-gut-brain axis. Dietary intervention, supplementation with suitable probiotics and fecal microbiota transplantation (FMT) can effectively alleviate or improve the symptoms of PD, and may be useful to treat the disease. However, due to the great heterogeneity of gut microbes, much work needs to be done to reveal the correlation between PD and intestinal microbiome.

To investigate the role of the prkd1 gene, a typical member of the protein kinase D (PRKD) family, in early heart development, zebrafish was used as an animal model in this study. Through prkd1 knockdown with morpholino antisense oligo (MO), it was found that knockdown of prkd1 resulted in abnormal cardiac de-velopment, such as pericardial edema, cyclization disorder, cardiac tube linearization. Zebrafish heart rate analysis indicated that, compared with the wild type, prkd1 knockdown zebrafish exhibited early arrhythmia. High-throughput transcriptome sequencing was performed for zebrafish embryos at 3 days post fertilization (dpf) in control and prkd1 knockdown groups. Differential gene enrichment analysis revealed that prkd1 knockdown was significantly associated with heart-related signaling pathways, such as the adrenergic signaling pathway in cardiomyocytes. Additionally, the results of high-throughput sequencing were confirmed by qRT-PCR, which showed that some genes associated with the cardiovascular system were markedly down-regulated. Meanwhile, identification of the essential factors in early heart development revealed that key cardiac regu-latory genes like tbx5a and gata4 were significantly down-regulated. These findings suggest that the prkd1 gene may affect early cardiac development by regulating cardiac development-related signal pathways and key genes, and play an important role at the early stage of heart development.

Novel coronavirus (SARS-CoV-2) infection increases the risk of myocardial injury, and the mole-cular mechanisms of pathogenesis remain to be elucidated. Herein, the changes of RNA editing in SARS-CoV-2 infected cardiomyocytes (infection group) and uninfected cardiomyocytes (mock group) were compa-red to explore the effects of the viral infection on cardiomyocytes from the perspective of RNA editing. RNA-seq data (GSE150392) of SARS-CoV-2 infection of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) was downloaded from the GEO database, and RNA editing sites (RESs) were iden-tified using SPRINT software. The changes in editing levels of cardiomyocytes in the infection and mock groups were compared, editing sites were annotated, and relevant functional analyses were performed. A total of about 92 899 base substitutions were detected, of which 87 670 were identified as A-to-I RESs, and 78 978 of these A-to-I editing sites were found to appear in the Alu regions. A-to-I editing sites tend to be dis-tributed in clusters, mainly in regions such as introns and intergenic regions. By screening the differential RESs between the infection and mock groups in A-to-I editing, it was found that 102 of the significantly differential RESs are up-regulated and 94 are down-regulated in editing level. These genes with significant differences in RES are involved in the biological processes of viral infection related GO (Gene Ontology), which are mainly enriched in viral process, virus life cycle, and defense response to virus, et al., and their Kyoto Encyclopedia of Genes and Genomes (KEGG) functional pathways are also mainly related to viral infec-tion. In contrast, the 11 screened genes with high-quality A-to-I RESs and significant differences in editing levels between groups are enriched in endocytosis, cytokine-cytokine receptor interaction, proteosome, nico-tinate and nicotinamide metabolism, and ferroptosis, et al.. The results showed that SARS-CoV-2 infection affects A-to-I RNA editing in cardiac myocytes, and the occurrence of such RNA editing events is a re-sponse of host cardiac myocytes to the viral infection.

In order to explore the distribution characteristics of major histocompatibility complex (MHC) classⅠchain-related (MIC) genes MICA and MICB in the Miao population of Guizhou to establish a genetic database that can be used to study the genetic characteristics and evolutionary history of certain diseases, PCR-sequence-specific primer (PCR-SSP) and PCR-sequence-based typing (PCR-SBT) methods were used to analyse the distribution of MICA and MICB genes from 150 Miao people in Guizhou Province by detecting their peripheral blood DNA samples. The differences in the distribution frequencies of MICA and MICB genes were analyzed and evaluated by the chi-square test. The genotyping results showed that the frequencies of MICA*019, MICA*045, MICA*008:04, MICA*002:01, MICA-A9, MICA-A4, MICB*005:02 and MICB*002 alleles were significantly higher than those of other alleles in the Miao population. Analysis of MICA-MICB gene sequence haplotypes revealed that the distribution frequency of MICA*019-MICB*005:02 and MI-CA*008:04-MICB*005:02 was higher, which were 17.40% and 16.30%, respectively, but only MICA*010-MICB*005:02, MICA*002:01-MICB*005:02 and MICA*009:01-MICB*005:02 were the MICA-MICB hap-lotypes with significant linkage disequilibrium (P<0.01). These results indicate that the MICA and MICB genes of Miao people in Guizhou are polymorphic and have significant linkage disequilibrium. The study provides experimental data for improving the MIC gene sequence database of ethnic minorities in China. These results could serve as the basis for future studies on the potential role of MICA and MICB in allogeneic organ transplantation and disease susceptibility in related ethnic groups.

With the development of biomedical technology, non-human primate (NHP) models become increa-singly widely used in basic scientific research. Meanwhile, many NHP species are on the verge of extinction due to habitat destruction, hunting and genetic isolation. Thus, it is important to improve the sperm cryop-reservation technology to conserve the genetic resources. This review mainly described the characteristics of NHP semen, introduced the methods of semen liquefaction and quality evaluation, and analyzed the effects of cryoprotectants, extenders and freezing methods on sperm cryopreservation. It also compared the relevant sperm parameters, discussed the puzzles and difficulties in sperm cryopreservation research, and proposed some feasible solutions. In conclusion, this paper reviewed the important research results of NHP sperm cryop-reservation in recent years, which may have certain reference value for development of NHP sperm cryopro-tectants and cryopreservation technology.

In order to explore the diversity of bacterial community structure and its influencing factors in the shallow sea area of the Haizhou Bay, both surface and bottom water samples from six sites in the bay were selected and analyzed based on high-throughput sequencing technology. The results showed that the abun-dance and diversity of bacterial communities in the bottom water samples were better than those in the sur-face water samples. There was difference in the evolutionary direction of bacterial communities between the bottom and surface water samples, but the difference was not significant between the two bacterial communi-ties. A total of 2 491 operational taxonomic units (OTUs) were detected in the Haizhou Bay, belonging to 32 phyla, 74 classes, 116 orders, 200 families and 364 genera. At the phylum level, the dominant bacteria in the bay were Proteobacteria, Bacteroidetes and Cyanobacteria. Ammonium (NH4-N) and chlorophyll (Chl) were found to be the main environmental factors affecting bacterial community structure. The study demonstrated that the diversity and distribution of bacterial community structure in the shallow sea area of the Haizhou Bay had a certain correlation with environmental factors, which provided a theoretical basis for the sustaina-ble development of the shallow sea ecosystem in this area.

The regulatory network constituted by transcription factors, enhancers, and microRNAs (miRNAs) is critical to the transcriptional regulation and progression of cancer. TEA domain transcription factor 1 (TEAD1) is a member of the TEA/ATTS domain family. However, it is unclear whether TEAD1, as a cancer-related transcription factor, is involved in the enhancer-miRNA network and the occurrence and develop-ment of hepatocellular carcinoma. Herein, 14 286 active enhancers that transcribe stable and unstable enhan-cer RNAs (eRNAs) were firstly identified by integrating CAGE-seq and GRO-seq data of HepG2 cell line, and confirmed to have previously reported histone modification characteristics (high H3K27ac signal and H3K4me1/H3K4me3 signal ratio). Subsequently, 2 550 TEAD1-binding enhancers (EnhTEAD1) were identified by in-tegrating 35 883 TEAD1-DNA binding sites obtained from ChIP-seq data. Compared with no TEAD1-binding enhancers (EnhnoTEAD1), the expression of active enhancer markers (H3K27ac, H3K4me1 and H3K4me3) and eRNAs on EnhTEAD1 was significantly increased. Furthermore, TEAD1 may act synergistically with four transcription factors (GATA4, HNF4A, YY1, and CTCF) to promote chromatin accessibility and spatial cyclization mediated by the EnhTEAD1 region. To study the regulatory network between EnhTEAD1 and miRNAs, small RNA sequencing was performed in HepG2 cells after TEAD1 was interfered using small in-terfering RNAs (siRNAs). A total of 68 differentially expressed miRNAs (EnhTEAD1-miRNAs) regulated by EnhTEAD1 were obtained by RNA-seq and Hi-C, and were found to be significantly involved in a variety of cancer-related biological processes and pathways. In summary, the study elucidates the regulatory mecha-nism of EnhTEAD1-miRNA network in hepatocellular carcinoma, and also provides a new potential target for treatment of hepatocellular carcinoma.

In order to understand the resources of amphibians and reptiles in Tongdao County, a total of 80 sample lines were set up in May, July and September of 2022 to carry out field investigation. A total of 24 species of amphibians belonging to 1 order, 6 families and 40 species of reptiles belonging to 2 orders, 12 families were recorded, accounting for 27.59% and 38.10% of the records in Hunan Province, respectively. It was found that the two types of animals in Tongdao County have obvious fauna characteristics: the Orien-tal species are dominated ones (67.19%), followed by widespread species (32.81%), and there are no Pa-laearctic species. Analysis of biodiversity showed that the Shannon-Wiener and Simpson indexes are 3.314 6 and 0.944 1, respectively, and the Pielou index is 0.797 0. The results of faunistic similarity showed that the faunistic division of the county is consistent with that of Hunan zoogeographic fauna. The survey made the current situation of amphibians and reptiles in the county more clearer, and would provide a scientific basis for protecting and studying the animal resources in this region.

To construct Huh-7 cell line stably expressing human angiotensin converting enzyme 2 (hACE2), the pWPXL-neo-hACE2 lentiviral vector without a fluorescent protein gene was constructed and co-trans-fected into HEK293T cells with psPAX2 and pMD2.G-VSVG to generate lentivirus. The obtained lentivirus particles were used to infect Huh-7 cells, and the cells that could stably express hACE2 protein were se-lected with hygromycin B. The expression of hACE2 in Huh-7 cells was detected by indirect immunofluo-rescence assay and Western-blot. Furthermore, the binding activity of Huh-7-hACE2 cells to the receptor-binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) pro-tein was analyzed by flow cytometry, and the susceptibility of Huh-7-hACE2 cells to SARS-CoV-2 was further determined with SARS-CoV-2 pseudovirus. The identified Huh-7-hACE2 cells were finally used in evaluating the neutralizing capacity of SARS-CoV-2 neutralizing antibody REGN10987 against pseudovirus. The results showed that the hACE2 protein was successfully expressed in Huh-7 cells, and the expressed hACE2 protein could bind to SARS-CoV-2 S protein RBD. Compared with Huh-7 cells, Huh-7-hACE2 cells were more susceptible to SARS-CoV-2 pseudovirus infection, and the neutralizing activity evaluated with pseudovirus was similar to that with SARS-CoV-2 reported in the literature. In conclusion, the study demonstrated that the Huh-7 cell line stably expressing hACE2 was successfully established, and could be used to evaluate the activity of corona virus disease 2019 (COVID-19) therapeutic monoclonal antibody. The established cell line is a useful tool to study the pathogenesis of SARS-CoV-2 and develop antiviral drugs and vaccines.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes corona virus disea-se 2019 (COVID-19), has been widely spread around the world for three years, and has spawned many va-riants. At present, the Omicron variant, which causes large-scale transmission in the world, has the charac-teristics of high infectivity and low toxicity, and has evolved into BA.4, BA.5, XBB and other subvariants. These strains can escape the vaccine-induced immunity, but neutralizing capacity elicited by vaccination can be restored to a certain degree by a booster dose. New vaccines and therapeutic drugs against the Omi-cron variant are available, providing better immunity and treatment methods for humans. This article sum-marized and analyzed the characteristics of Omicron strains, human immune mechanism against the infec-tion, current vaccine and drug development, and COVID-19 sequelae. It also made suggestions on how to cope with SARS-CoV-2 in the future.

Abstract: In order to identify the hub genes and biomarkers of hepatocellular carcinoma (HCC) by bioinfor-matics methods, the GSE112790 dataset from the Gene Expression Omnibus (GEO) and HCC data from The Cancer Genome Atlas (TCGA) database were screened for differential expression analysis. A total of 151 dif-ferentially expressed genes (DEGs) were identified and 10 hub genes were screened. Survival analysis indi-cated that upregulated expression of BUB1B, CDC20, ASPM and DLGAP5 genes significantly affected the prognosis of patients. Analysis of Oncomine database showed that expression levels of BUB1B, CDC20 and DLGAP5 were obviously related to tumor grade and vascular invasion. From HPA database and immunohis-tochemistry of liver cancer tissue microarray, it was found that expression levels of CDC20 and DLGAP5 in HCC tissues were higher than those in normal tissues. Cox analysis showed that the high expression of CDC20 and DLGAP5 could be used as an independent risk factor for the prognosis of patients with HCC. Moreover, the CDC20 methylation level was also found to be an important factor affecting its expression level and a va-riety of immune cell contents. In conclusion, CDC20 may be a potential biomarker for HCC prognosis evalua-tion and therapeutic target.