BACKGROUND:The self-renewal and multi-directional differentiation of pluripotent stem cells possess the potential to revolutionize people's understanding of biology,medicine,development,and disease.Stem cells play an important role in the early stage of embryonic development,and the study of them could be beneficial to understanding of the basic principles of biological development and tissue or organ formation,exploring the potential mechanisms of various diseases,studying the repair and regeneration of damaged tissues or organs,and promoting drug discovery and personalized treatment. OBJECTIVE:To review the research progress of pluripotent stem cells,summarize and categorize the fundamental types of pluripotent stem cells,and elucidate the lineage situations of various types of pluripotent stem cells in common mammals. METHODS:PubMed,Web of Science,CNKI,and WanFang databases were searched systematically,with the keywords"pluripotent stem cells;embryonic stem cells;induced pluripotent stem cells;expanded potential stem cells;livestock pluripotent stem cells"in English and Chinese.The 99 articles related to mammalian pluripotent stem cells were systematically screened according to inclusion and exclusion criteria,and then reviewed. RESULTS AND CONCLUSION:(1)According to classical theory in mouse embryonic stem cell research,the pluripotent state of stem cells is divided into two forms:na?ve and primed.Na?ve state corresponds to the inner cell mass of pre-implantation embryos before attachment to the uterine wall,while primed state corresponds to the epiblast after implantation.These two states exhibit significant differences in epigenetic features,transcriptional activity,external signal dependency,and metabolic phenotype.It is later discovered that there is an intermediate state between na?ve and primed called formative pluripotency.Therefore,the pluripotency of pluripotent stem cells is a continuous developmental process rather than a unique cell state.(2)In addition to obtaining pluripotent stem cells from the inner cell mass,there are various methods and lineages for acquiring pluripotent stem cells,including embryonic germ cells established using primitive germ cells from mouse embryos,induced pluripotent stem cells created by the dedifferentiation of adult mouse and human fibroblasts with four factors—Oct3/4,Sox2,c-Myc,and Klf4;embryonic stem cell-like cell lines cultured from somatic cell nuclear transfer,parthenogenesis,neonatal or adult testicular or ovarian tissue,very small embryonic-like stem cells derived from various adult tissues and expanded pluripotent stem cells derived from pre-implantation stages.These pluripotent stem cells all share the common characteristics of continuous self-renewal,expressing core pluripotency factors and possessing the ability to differentiate into the three primary germ layers.(3)Currently,pluripotent stem cells are being used for disease modeling to study the mechanisms of various diseases and develop new drugs.Simultaneously,scientists are attempting to use pluripotent stem cells to cultivate various tissues and organs,offering new possibilities for regenerative medicine and transplantation.However,the clinical application of pluripotent stem cells faces safety challenges,including issues of cell mutations and immune rejection.Continual improvement in the methods of generating pluripotent stem cells will make them safer and more efficient for clinical applications.(4)Based on the methods of obtaining and lineage establishment of pluripotent stem cells in mice and humans,various types of pluripotent stem cells have been established in livestock,including embryonic stem cells,induced pluripotent stem cells,germ lineages of pluripotent stem cells,and expanded potential stem cells.Research on livestock pluripotent stem cells opens up new avenues for animal reproduction,breeding,genetic engineering,disease modeling,drug screening,and the conservation of endangered wildlife.

In 1929, George Soulié de Morant and Paul Ferreyrolles co-authored their first acupuncture-moxibustion paper titled "L' Acuponcture en Chine vingt siècles avant J.-C. et la réflexothérapie moderne", greatly advancing the development of acupuncture-moxibustion in Europe. Their paper systematically explains the holistic view and the concept of yin-yang balance in traditional Chinese medicine, describes the techniques of acupuncture and moxibustion, innovatively classifies acupuncture-moxibustion as "reflexotherapy", organizes the effects of certain acupuncture points illustrated on human acupoint atlas; and for the first time, it summarizes the correspondence between acupuncture points and Weihe trigger points. In the historical background of the neo-Hippocratic movement, they used the existing theories at that time to explain acupuncture, and adopted the analogical medicine to explore the mechanisms of acupuncture-moxibustion, which gradually initiated the modern era of acupuncture-moxibustion in France. Such research method is conducive to reducing the unfamiliarity of acupuncture-moxibustion among westerners, deepening their understanding of its theories and therapeutic effect, and also integrating it with other medical research. It breaks through the limitations of traditional theories and obtains the self-improvement and progress.
Humans ; Moxibustion/history* ; Acupuncture Therapy/history* ; China ; History, Ancient ; History, 20th Century ; Acupuncture/history* ; Reflexotherapy/history* ; Acupuncture Points ; History, 19th Century ; Medicine, Chinese Traditional/history*

Celastrol, a pentacyclic triterpenoid compound derived from the root of Chinese herb Tripterygium wilfordii Hook.f.,can inhibit the growth of various types of malignant tumors. However, it still has some limitations, including high toxicity, poor water solubility, and low targeting efficiency. Therefore, structural modification of celastrol has become a research hotspot in recent years. The structural modifications of celstrol reported have focused on C-20-COOH and C-2, C-3, C-6 or multiple sites of AB ring. This review provides an overview of the research progress of anti-tumor celastrol derivatives in recent years according to different structural modification sites and purposes, such as enhancing the inhibitory effect on the Hsp90-Cdc37 protein-protein interaction, and modification methods, including principles of parallelism and targeting specific sites. In addition, it briefly discusses the antitumor activity, mechanism of action, and structure-activity relationship of these derivatives, aiming to provide theoretical guidance for the discovery of new celastrol derivatives with high efficiency, low toxicity, and strong selectivity.

Objective To investigate the distribution of B cells in both tumor and non-tumor tissues of gastric cancer patients,analyze their phenotypic characteristics and explore the impact on T cell proliferation.Methods Immunohistochemical staining was utilized to detect the expression of B cell surface marker CD 19 in tumor and non-tumor tissues from 33 gastric cancer patients.The expression levels of chemokine receptors and immunoglobulin molecules on B cells in both tumor and non-tumor tissues were measured using flow cytometry.Chemotaxis experiments were conducted to examine the role of the CXCL12-CXCR4 axis in B cell chemotaxis.B cells isolated and purified from both tissue types were co-cultured with autologous peripheral T cells to assess their effect on T cell proliferation.Results There were significantly more B cells infiltrated in tumor tissues than those infitrated in the non-tumor tissues of gastric cancer patients(P＜0.01),and CXCR4 was highly expressed on tumor-infiltrating B cells compared with B cells derived from non-tumor tissues(P＜0.05).The Cancer Genome Atlas(TCGA)analysis indicated that the expression level of CXCL12 in tumor tissues was positively correlated with the expression level of CD19 in gastric cancer patients(r=0.15,P＜0.01).And the expression level of CXCL12 in tumor tissues of the gastric cancer patients was also positively correlated with the number of B cells infiltrated in tumor tissues.Chemotaxis experiments confirmed that the CXCL12-CXCR4 axis was involved in promoting B cell chemotaxis(P＜0.05).Although B cells in tumor and non-tumor tissues had similar levels of IgM,IgG,and IgA expression,tumor-infiltrating B cells significantly inhibited the proliferation of T cells when compared with B cells derived from non-tumor tissues(P＜0.01).Conclusion There are more B cells infiltrated in gastric cancer tissues,which may be recruited to tumor tissues through the CXCL12-CXCR4 axis,and then inhibit T cell proliferation to promote the progression of gastric cancer.

Depression is a psychosomatic disorder.The rising incidence rate of depression in recent years is placing a heavy economic burden on societies around the world.Morinda officinalis oligosaccharides(MOOs)are active substances extracted from the Chinese herb Morinda officinalis that can soothe depression and calm the mind,tonify the kidneys,and benefit the intellect,as well as improve cognitive disorders in patients to a certain extent.On the basis of the hypothesised pathological mechanism of depression,this study explains the link between MOOs and depression by reviewing existing studies.We propose that MOOs can improve depression through mechanisms that regulate the levels of monoamine neurotransmitters,enhance neuroplasticity,regulate the function of the HPA axis and levels of cytokines,and influence gut microbiota.This paper provides new ideas for research on the antidepressant effects of MOOs.

Objectives To construct a prediction model of relapse in diffuse large B-cell lymphoma within two years after complete remission based on multiple randomized empirical kernel learning machine to provide a basis for patient treatment decisions.Methods Using the information of 445 patients who met the requirements of this study in the electronic medical record database of a tertiary hospital in Shanxi Province from 2010 to 2020,a relapse prediction model was constructed based on five common categories of imbalance treatment methods and a multiple stochastic empirical kernel learning machine,and compared with the five classifiers.Results The recurrence prediction model based on SMOTE Tomek Links+multiple randomized empirical kernel learning machine achieved optimal classification performance(accuracy=0.89,precision=0.87,recall=0.92,f1-Score=0.89,brier score=0.11).Conclusion For the actual DLBCL dataset,in this paper,we used SMOTE Tomek links to process the imbalance data and construct a multiple randomized empirical kernel learning machine,which achieves the optimal model performance with low computational complexity and can provide a powerful reference for DLBCL recurrence prediction.

Objective:To construct pancreatic cancer cell lines expressing luciferase and mesothelin (MSLN), and evaluate the feasibility of using them as target cells in analyzing the cytotoxicity activity of immune cells.Methods:Lentiviral vectors expressing luciferase and MSLN genes were constructed, and pancreatic cancer cell lines were infected after lentivirus packaging. Single-cell clones were obtained by limited dilution following antibiotic screening, and the stable expression of the target genes were verified. These cells were used as target cells to detect the cytotoxicity of immune cells by real-time cell analysis (RTCA) and luciferase activity. Besides, these luciferase-expressing cells were transplanted into B-NDG mice to establish the animal models of pancreatic cancer, and in vivo optical imaging technology was used to detect the expression of luciferase and monitor the tumor growth in mice. The cytotoxicity of chimeric antigen receptor T (CAR-T) cells was verified in these animal models. Results:Three pancreatic cancer cell lines, panc-1-luc, panc-1-luc-MSLN and capan-2-luc, that could stably express luciferase and MSLN genes were successfully constructed. The expression of the reporter gene in these cells were high, and positively correlated with the number of cells. There were 95.6% of panc-1-luc-MSLN cells expressing MSLN. MSLN-CAR-T cells had specific killing effect on MSLN-positive panc-1-luc-MSLN cells and capan-2-luc cells, with the minimum killing rates of (70.00±18.19)% and (57.00±5.29)%, respectively. But they had no cytotoxicity to MSLN-negative panc-1-luc cells. RTCA results showed that MSLN-CAR-T cells were able to lyse all three pancreatic cancer cell lines, and the minimum killing rates were (56.33±7.64)%, (93.00±2.65)% and (26.33±28.15)%, respectively. The killing of target cells by NK-92MI cells was not depended on MSLN expression. The cytotoxicity in the mice models of pancreatic cancer was consistent with the results in vitro. The in vivo and in vitro test results suggested that the expression of luciferase by target cells could reflect the cytotoxicity of immune cells. Conclusions:This study establishes three pancreatic cancer cell lines stably expressing luciferase, which can be used to evaluate the cytotoxicity of immunotherapy products targeting tumor cells in vitro and in vivo.

Background Gastric carcinogenesis is a multifactorial and complex process, in which long non-coding RNAs (lncRNAs) play important roles as oncogenes or antioncogenes. Research has found that the expression of lncRNA LINC02859 is down-regulated in gastric cancer tissues and correlated with the degree of tumor differentiation and TNM stage, and also plays an important role in the development of malignant transformation of cells induced by environmental carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), but its mechanism of action is still unclear. Objective To explore the role and potential regulatory mechanism of gastric cancer-associated lncRNA LINC02859 in MNNG-induced malignant transformation of human normal gastric mucosal cells (GES-1). Methods A total of 110 gastric cancer patients from a high incidence area of gastric cancer in Gansu Province were selected, and their cancer tissues and normal gastric mucosa tissues adjacent to the cancer were collected to detect the expression level of LINC02859 by real-time quantitative PCR (RT-qPCR). High-throughput sequencing and bioinformatics analysis of the tissues were used to identify the potential signaling pathways regulated by the genes co-expressed with LINC02859. GES-1 cells at 70%-80% cell fusion with low cell passage number and normal morphology were incubated with 0, 0.25 and 0.5 μmol·L⁻¹ MNNG solution for 48 h and the LINC02859 expression level was detected. Cell proliferation activity was detected by Cell Counting Kit-8 (CCK-8), clone formation was detected by plate clone formation assay, and cell migration ability was detected by scratch assay to evaluate the effects of MNNG on cell morphology and function. The expression levels of key proteins of Wnt signaling pathway were detected by RT-qPCR and Western blotting. Results The RT-qPCR results showed that LINC02859 was lowly expressed in the gastric cancer tissues compared with the paracancerous tissues, and the difference was statistically significant (P < 0.05). The pathway enrichment analysis showed that LINC02859 potentially regulated the Wnt pathway. The in vitro malignant transformation assay suggested that after the MNNG exposure, the malignant cells of passage 5 (MC-5) had altered morphology, increased number of colony formation, and higher proliferation and migration ability than the control cells; compared with the normal GES-1 cells, LINC02859 gene expression levels were reduced in the 0.25 μmol·L⁻¹ and the 0.5 μmol·L⁻¹ MNNG-exposed GES-1 cells; the expression levels of key proteins of the Wnt pathway, transcription factor 7 (TCF7), Axis inhibitor (Axin1), phosphorylation of glycogen synthase kinase-3 beta (p-GSK-3β), casein kinase 1 (CK1), and β-catenin, were elevated in the cells after 0.5 μmol·L⁻¹ MNNG exposure (P < 0.05); whereas, overexpression of LINC02859 suppressed the activating effect of MNNG on the Wnt pathway. Conclusion LINC02859 is lowly expressed in the cancer tissues of gastric cancer patients. MNNG exposure induces morphological and functional changes in GES-1 cells, down-regulated expression of LINC02859, and activation of the Wnt signaling pathway; overexpression of LINC02859 inhibits the activation of the Wnt signaling pathway in the gastric carcinogenesis induced by MNNG exposure.

Objective:To investigate the clinical characteristics and prognostic factors of TCF3-PBX1 fusion gene-positive childhood B-cell precursor acute lymphoblastic leukemia (B-ALL).Methods:The clinical data of 1 287 newly diagnosed children with B-ALL who were admitted to five hospital in Fujian province (Fujian Medical University Union Hospital, the First Affiliated Hospital of Xiamen University, Zhangzhou Affiliated Hospital of Fujian Medical University, Quanzhou First Hospital Affiliated to Fujian Medical University, Nanping First Hospital of Fujian Province) from April 2011 to December 2020 were retrospectively analyzed. According to the results of TCF3-PBX1 fusion gene testing, all the patients were divided into TCF3-PBX1-positive group and TCF3-PBX1-negative group. The clinical characteristics, early treatment response [minimal residual disease (MRD) at middle stage and end of induction chemotherapy] and long-term efficacy [overall survival (OS) and event-free survival (EFS)] of the patients in both groups were compared. Kaplan-Meier method was used for survival analysis. The prognostic factors of TCF3-PBX1-positive B-ALL were analyzed by using Cox proportional hazards model. Among 83 children with TCF3-PBX1-positive B-ALL, the treatment regimens, risk stratification and efficacy evaluation of 62 cases were performed by using Chinese Children's Leukemia Group (CCLG)-ALL 2008 regimen and 21 cases were performed by using Chinese Children's Cancer Group (CCCG)-ALL 2015 regimen, and the efficacy and incidence of serious adverse events (SAE) between the two groups compared.Results:Among 1 287 B-ALL patients, 83 patients (6.4%) were TCF3-PBX1-positive. The proportion of patients with initial white blood cell count (WBC)≥50×10 9/L in the TCF3-PBX1-positive group was higher than that in the TCF3-PBX1-negative group, while the proportions of patients with MRD ≥1% on induction chemotherapy day 15 or day 19, and MRD ≥0.01% on induction chemotherapy day 33 or day 46 in the TCF3-PBX1-positive group were lower than those in the TCF3-PBX1-negative group (all P < 0.05). Univariate Cox regression analysis showed that MRD ≥1% on induction chemotherapy day 15 or day 19 and TCF3-PBX1 ≥0.01% on induction chemotherapy day 33 or day 46 were risk factors for OS and EFS (all P < 0.05). Multivariate analysis showed that MRD ≥1% on induction chemotherapy day 15 or day 19 was an independent risk factor for OS ( HR = 10.589, 95% CI 1.903-58.933, P = 0.007) and EFS ( HR = 10.218, 95% CI 2.429-42.980, P = 0.002). TCF3-PBX1≥0.01% on induction chemotherapy day 33 or day 46 was an independent risk factor for EFS ( HR = 6.058, 95% CI 1.463-25.087, P = 0.013) but not for OS ( HR = 3.550, 95% CI 0.736-17.121, P = 0.115). The 10-year EFS and OS rates of the TCF3-PBX1-positive group were 84.6% (95% CI 76.9%-93.1%) and 89.1% (95% CI 82.1%-96.6%), and the differences between the two groups were not statistically significant (both P > 0.05). Among 80 children who received standardized treatment, compared with children who were treated with CCLG-ALL 2008 regimen, the incidence of infection-related SAE was lower in children who were treated with CCCG-ALL 2015 regimen [0 (0/21) vs. 20.3% (12/59), χ2 = 5.22, P = 0.022], but there were no statistical differences in treatment-related mortality, relapse rate, EFS and OS between the two groups (all P > 0.05). Conclusions:Children with TCF3-PBX1-positive B-ALL have a good prognosis, and MRD≥1% at middle stage of induction chemotherapy and TCF3-PBX1≥0.01% at the end of induction chemotherapy may be influencing factors for poor prognosis. CCCG-ALL 2015 regimen can reduce infection-related SAE while achieving good efficacy.

Objective:To explore the application and practice of "flipped classroom" in the teaching of general surgery interns.Methods:A total of 20 internship groups (3 to 5 people in each group) were randomly selected from the general surgery practice group in the Department of General Surgery of the Second Clinical Medical College of North Sichuan Medical College. They were randomly divided into the flipped group (45 people) and the traditional group (40 people), with 10 subgroups in each group. The flipped group adopted the flipped classroom teaching mode (students' self-study by handing out materials before class, students and teachers' discussion in class, and students and teachers' evaluation after class), while the control group adopted the current conventional teaching mode (students' preview before class, teachers' explanation in class, and teachers' question answering after class). At the end of the teaching, a questionnaire was used to evaluate the participation and completion of each student. The teaching effect was evaluated by medical history collection and case analysis. The participation, completion, and teaching effect between the two groups were compared and analyzed. SPSS 23.0 software was used for t-test and Chi-square test. Results:The participation of the flipped group was better than that of the traditional group [(17.45±1.83) vs. (15.57±1.52)], and the difference was statistically significant ( P < 0.05). There was no statistically significant difference between the flipped group and the traditional group. There was no significant difference in medical history collection scores between the two groups. The case analysis of the flipped group was better than that of the traditional group [(87.30±6.06) vs. (81.50±5.88), P < 0.05]. The questionnaire shows that about 90% of the students think that flipped classroom can improve their interest in learning [96% (43/45)], improve their autonomous learning ability [89% (40/45)], and have better learning effect. At the same time, 78% (35/45) of students think that learning time is too long. Conclusion:The flipped classroom teaching model can improve the teaching participation of general surgery students, improve students' interest in learning, improve their self-learning ability, and improve students' thinking ability of medical record analysis.