ObjectiveTo explore the potential categories and characteristics of the public hospice care demand in Hainan Province， and analyze different potential types of influencing factors， so as to provide reference for relevant departments to improve the public awareness and demand of hospice care. MethodsUsing convenience sampling method， select 6484 cities of the public as the survey object， using the general data questionnaire， the hospice care demand questionnaire of the potential profile analysis， and analyze the influencing factors of the public hospice care demand category. ResultsThe characteristics of the hospice care demand in Hainan Province were divided into three potential categories： low demand group （14.19%）， medium demand group （49.99%） and high demand group （35.82%）. Multivariate analysis showed that gender， age， education level， cultural belief， and life-death education experience were the main influencing factors of public hospice care demand （p<0.05）. Males， those aged 41-60 years， and those with high school education or below had relatively lower hospice care demand， while those with life-death education experience had relatively higher demand. ConclusionRelevant departments should focus on hospice care knowledge popularization and demand enhancement for males， middle-aged groups， and people with low education levels， while strengthening universal life-death education through stratified and classified publicity strategies and educational interventions to improve different populations’ awareness and acceptance of hospice care.

Objective:To construct a prognostic model for ovarian cancer based on programmed cell death receptor-1(PD-1)and B lymphocyte infiltration.Methods:A total of 102 patients with ovarian cancer admitted to the First Affiliated Hospital of Gannan Medical College from February 2019 to February 2022 were selected.PD-1 expression and CD20+B cell infiltration degree in cancer tissues were detected,relationship between them and pathological characteristics were analyzed,and correlation between PD-1 expres-sion with B lymphocyte infiltration degree and survival rate of patients were analyzed.Independent influencing factors of poor prognosis of ovarian cancer patients were analyzed,prediction model was built and validated according to independent factors.Results:PD-1 expression and CD20+B cell infiltration degree were closely related to tumor size,FIGO stage,tumor differentiation and lymph node metastasis(P<0.05).PD-1 expression was negatively correlated with B-lymphocyte infiltration degree(P<0.05).Patients with high PD-1 expression and low B-lymphocyte infiltration had higher mortality(P<0.05).FIGO stage,lymph node metastasis,PD-1 expres-sion and CD20+B cell infiltration were independent factors affecting poor prognosis of ovarian cancer patients.Prediction model was constructed by independent influencing factors,which had good differentiation and accuracy.Conclusion:PD-1 expression is closely related to CD20+B cell infiltration degree.There is a close relationship between PD-1 and CD20+B cell infiltration with clinico-pathological characteristics and prognosis of ovarian cancer patients,which are independent influencing factors for poor prognosis of ovarian cancer patients.

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear. Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP). Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin. Conclusions PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

OBJECTIVES: Wild-caught Microtus fortis (M. fortis) at the age of 9-15 months can develop epithelial ovarian cancers similar to human epithelial ovarian cancers under natural conditions during experimental animal breeding, but its pathological types and biological characteristics remain unclear. This study aims to analyze the biological characteristics of spontaneous ovarian cancer in M. fortis, intending to develop M. fortis as an animal model for human epithelial ovarian cancer. METHODS: The female M. fortis (9-15 months old) with spontaneous ovarian cancer were selected as the experimental group, and healthy M. fortis from the same litter were selected as the control group. The ovarian pathological changes of the two groups were observed by dissection. Blood routine and biochemical indicators were measured by biochemical analysis. Hematoxylin and eosin (HE) staining was performed to observe the pathological changes in the ovarian cancer tissue of M. fortis. Immunohistochemical staining was used to detect the protein expression of common ovarian cancer markers, and real-time RT-PCR was used to analyze the transcription levels of ovarian cancer-related genes. RESULTS: Spontaneous ovarian cancer in M. fortis mainly affects both ovaries, with tumors appearing solid or cystic. HE staining and histopathological analysis confirmed that the ovarian tumors originated from ovarian surface epithelium. Compared to the control group, the experimental group showed significantly decreased hemoglobin (P<0.01), hematocrit (P<0.05), albumin (P<0.05), and blood glucose levels (P<0.01), while lymphocyte percentage (P<0.05), monocyte percentage (P<0.05), cholesterol (P<0.01), and progesterone (P<0.01) levels were significantly increased. Expression of ovarian cancer-related genes, including ID3, CDC42, RHOA, RB1CC1, NF1, PIN1, MIB1, PDS5A, MCM7, and MLH1, was significantly downregulated (all P<0.05), while PAX8 gene expression was significantly upregulated (P<0.05). Immunohistochemical results showed that Wilms' tumor gene 1 (WT1) protein was mainly distributed throughout the cell, with significantly higher expression in ovarian cancer M. fortis. Tumor protein 53 (TP53) was expressed in both healthy and ovarian cancer M. fortis and was distributed throughout the cell. Hepatocyte nuclear factor 1 beta (HNF1B) and progesterone receptor (PR) protein were highly expressed in the ovarian tissue of healthy M. fortis but were significantly reduced in the ovarian cancer M. fortis, though both were located in the cytoplasm. CONCLUSIONS Spontaneous ovarian cancer in M. fortis is serous ovarian cancer. Compared to healthy M. fortis, significant differences were observed in ovarian tissue morphology, biochemical indicators, ovarian cancer-related gene expression, and protein expression, which show similarity to the biological characteristics of human serous ovarian cancer. This suggests that M. fortis could be an ideal animal model for studying human serous ovarian cancer.
Female ; Ovarian Neoplasms/metabolism* ; Animals ; Carcinoma, Ovarian Epithelial ; Disease Models, Animal ; Humans ; Neoplasms, Glandular and Epithelial/metabolism* ; Ovary/pathology*

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear. Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP). Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin. Conclusions PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear. Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP). Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin. Conclusions PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

Objective:To explore the clinical efficacy and safety of a multimodal treatment regimen integrating radiotherapy, chemotherapy, and immunotherapy in patients with human epidermal growth factor receptor 2 (HER2) -negative locally advanced or advanced gastric cancer.Methods:A total of 34 patients with unresectable, HER2-negative, locally advanced or metastatic gastric/gastroesophageal junction (G/GEJ) adenocarcinoma admitted to the Affiliated Cancer Hospital of Guizhou Medical University from September 2021 to March 2024 were selected as study objects. Participants received one cycle of either XELOX regimen (capecitabine + oxaliplatin) or SOX regimen (S-1 + oxaliplatin) with immunotherapy (sintilimab or nivolumab) . The process was succeeded by radiotherapy targeted at the primary G/GEJ tumor and regional lymph nodes. In selected cases, sequential radiotherapy was also administered for distant metastases. The primary endpoint was objective response rate (ORR) , and secondary endpoints were disease control rate (DCR) , clinical symptom response, changes in Karnofsky performance status (KPS) score, progression-free survival (PFS) , and adverse reactions. Clinical efficacy was assessed in accordance with Response Evaluation Criteria in Solid Tumors version 1.1. Adverse reactions were assessed and graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events version 5.0 and the Chinese Society of Clinical Oncology guidelines for management of immune checkpoint inhibitor-related toxicity. With a median follow-up of 7 months (range: 2.3 to 30 months) , the final evaluation considered the best response documented throughout follow-up. Survival curves were constructed utilizing Kaplan-Meier analysis.Results:By the end of follow-up, an overall ORR of 58.8% (20/34) and DCR of 70.6% (24/34) were observed. The ORR of lesions by radiotherapy reached 73.8% (48/65) and the DCR reached 92.3% (60/65) . Univariate analysis showed that the ORR of female patients (84.6%, 11/13) was higher than that of male patients (42.9%, 9/21) , and the ORR of patients with distant lymph node metastasis alone (83.3%, 15/18) was higher than that of patients with distant lymph node metastasis combined with organ metastasis or organ metastasis alone (18.2%, 2/11) , with statistically significant differences ( P=0.030; P=0.010) . There were no statistically significant differences in ORR among patients with different age ( P=0.487) , KPS score ( P=0.198) , primary tumor location ( P=0.280) , histological differentiation ( P=0.668) , chemotherapy regimen ( P=0.728) , or immunotherapy regimen ( P>0.999) . Twenty-two of 23 (95.7%) patients with upper abdominal pain were relieved, 10 of 21 (47.6%) patients with appetite loss were relieved, 15 of 17 patients with upper abdominal distension were relieved, 13 of 14 patients with melena were relieved, 6 of 7 patients with eating obstruction were relieved, 3 of 4 patients with metastatic site pain were relieved, and 2 patients with hematemesis were relieved. KPS score enhanced in 82.4% (28/34) of patients, remained stable in 11.8% (4/34) , and declined in 5.8% (2/34) . The median PFS of the 34 patients was 7.9 months. The most common adverse reactions during radiotherapy combined with chemotherapy and immunotherapy were hematological adverse reactions, in which neutropenia accounted for the highest proportion (91.2%, 31/34) , followed by anemia (50.0%, 17/34) . Fatigue was the most common non-hematological adverse reaction (50.0%, 17/34) , followed by nausea and vomiting (26.5%, 9/34) . The adverse reactions of 6 patients receiving immune monotherapy maintenance were anemia, hypothyroidism, transaminase elevation, proteinuria, fatigue, and rash, all of which were grade 1-2. Conclusions:Radiotherapy combined with chemotherapy and immunotherapy shows good short-term clinical efficacy in patients with HER2-negative locally advanced or advanced gastric cancer, and the overall adverse reactions are tolerable. Female or patients with distant lymph node metastasis alone may be the preferred population for this study protocol.

Objective:To construct a prognostic model for ovarian cancer based on programmed cell death receptor-1(PD-1)and B lymphocyte infiltration.Methods:A total of 102 patients with ovarian cancer admitted to the First Affiliated Hospital of Gannan Medical College from February 2019 to February 2022 were selected.PD-1 expression and CD20+B cell infiltration degree in cancer tissues were detected,relationship between them and pathological characteristics were analyzed,and correlation between PD-1 expres-sion with B lymphocyte infiltration degree and survival rate of patients were analyzed.Independent influencing factors of poor prognosis of ovarian cancer patients were analyzed,prediction model was built and validated according to independent factors.Results:PD-1 expression and CD20+B cell infiltration degree were closely related to tumor size,FIGO stage,tumor differentiation and lymph node metastasis(P<0.05).PD-1 expression was negatively correlated with B-lymphocyte infiltration degree(P<0.05).Patients with high PD-1 expression and low B-lymphocyte infiltration had higher mortality(P<0.05).FIGO stage,lymph node metastasis,PD-1 expres-sion and CD20+B cell infiltration were independent factors affecting poor prognosis of ovarian cancer patients.Prediction model was constructed by independent influencing factors,which had good differentiation and accuracy.Conclusion:PD-1 expression is closely related to CD20+B cell infiltration degree.There is a close relationship between PD-1 and CD20+B cell infiltration with clinico-pathological characteristics and prognosis of ovarian cancer patients,which are independent influencing factors for poor prognosis of ovarian cancer patients.

Objective:We genetically modified our non-replicating vaccinia virus NTV to improve its immunogenicity.Methods:We constructed NTV-modified strain NTV-ΔF1L-C7L by homologous recombination of vaccinia virus based on CRISPR-Cas9 technology by inserting the C7L gene while deleting the F1L gene. The recombinant virus NTV-ΔF1L-C7L was then immunized with 10 7 PFU in BALB/c mice, and the levels of humoral and cellular immunity induced by NTV-ΔF1L-C7L were detected by ELISA and ELISpot method, respectively, and the levels of neutralizing antibodies were determined by the phage-reduced neutralization assay. Results:The PCR and western- blot identification proved that the F1L gene of the constructed NTV-modified strain NTV-ΔF1L-C7L was missing, while the C7L gene was inserted back in the region, and the C7L gene could be expressed normally, indicating that the recombinant virus was constructed correctly. After immunization of mice with NTV-ΔF1L-C7L, ELISA result showed that the recombinant virus NTV-ΔF1L-C7L induced a higher level of IgG antibody than NTV; ELISpot result also showed that the recombinant virus was able to induce a higher level of IFN-γ; and the result of plaque reduction neutralization test showed that the recombinant virus was able to induce a higher level of IFN-γ antibody than that of NTV.Conclusions:We correctly constructed the NTV gene-modified strain NTV-ΔF1L-C7L, which induced stronger humoral and cellular immunity compared with NTV, and provided reference data for the research and development of replacement products for smallpox or monkeypox vaccines.

BACKGROUND:The research of dental stem cells in the fields of regenerative medicine and tissue engineering has been deepening,bringing hope for the repair of tooth-related tissues and the treatment of systemic diseases.However,there is a lack of systematic research and analysis on the biological characteristics of dental stem cells in different age groups. OBJECTIVE:To explore the biological characteristics of the human deciduous tooth and permanent tooth pulp stem cells cultured in umbilical cord blood platelet lysate to provide a reliable basis for human platelet lysates to replace fetal bovine serum. METHODS:The pulp tissues of deciduous teeth,juvenile permanent teeth and adult permanent teeth were taken out and cultured in DMEM/F-12 medium supplemented with 10%fetal bovine serum or different concentrations(5%,10%and 15%)of human platelet lysates.Cell proliferation in the four groups was detected by cytometry.The optimal concentration of human platelet lysates was selected for subsequent experiments.Under the optimal concentration of human platelet lysates,human deciduous tooth and juvenile and adult permanent tooth pulp stem cells were cultured in vitro.The cell growth status was observed under the microscope.The specific antigen on the cell surface was detected by flow cytometry.The cell proliferation ability was tested by the cell counting method and CCK-8 assay.The cell differentiation ability in vitro was observed by a three-line differentiation assay. RESULTS AND CONCLUSION:(1)The cell proliferation rate of the 10%human platelet lysate group was the highest.(2)In all three groups,fusiform fibrous cells grew and expanded from around the tissue block.There was no significant difference between deciduous teeth and juvenile permanent tooth cells,but the adult permanent tooth cells were larger than the deciduous and juvenile permanent tooth cells of the same generation.(3)The results of flow cytometry showed that deciduous teeth,juvenile permanent teeth and adult permanent teeth conformed to the phenotypic characteristics of mesenchymal stem cells.(4)The proliferative capacity of adult permanent dental pulp stem cells was significantly lower than those of deciduous teeth and juvenile permanent dental pulp stem cells(P<0.01).(5)mRNA expressions of osteoblast-related genes alkaline phosphatase and bone morphogenetic protein 2,lipoprotein lipase and peroxisome proliferator-activated receptor γ2,mRNA expressions of chondroblast related gene type II collagen α1 and cartilage oligomeric matrix protein in adult pulp stem cells of permanent teeth were significantly lower than those of deciduous teeth and juvenile permanent teeth pulp stem cells(P<0.01).(6)Compared with adult dental pulp stem cells,human deciduous teeth and juvenile permanent teeth dental pulp stem cells have the stronger proliferative capacity and multidirectional differentiation potential,and are more suitable for clinical research and disease treatment.