Objective:To separate and purify the polysaccharides from Polygonatum filipes,characterize their primary structure and investigate the immunomodulatory effects on RAW264.7 macrophages.Methods:Crude polysaccharides from Polygonatum filipes were extracted by ultrasound assisted method,then Polygonatum filipes polysaccharides(CSPFPs)were obtained after elimination of the proteins with combined papain-Sevag method.The total sugar content was determined by phenol-sulfuric acid method.Structures of CSPFPs were analyzed by fourier transform infrared spectroscopy(FT-IR),high performance gel permeation chromatography(HPGPC)and high performance liquid chromatography(HPLC).Effects of CSPFPs on cell viability,pinocytic activity,TNF-α secretion,MAPK and NF-κB signaling pathways of RAW264.7 cells were explored by MTT,Neutral red,ELISA and Western blot,respectively.Results:Extraction rate of CSPFPs by ultrasound-assisted method was 41.61%,which contained total sugar content of 94.00%.CSPFPs with Mw of 3 125 Da was composed of arabinose(1.85%),galactose(6.14%),glucose(56.41%)and mannose(35.60%).The in vitro experiments showed that CSPFPs were non-cytotoxic and enhanced the pinocytic activity,TNF-α secretion and phosphorylation levels of p38,ERK,JNK,p65,IκB and IKK,indicating the activation of MAPK and NF-κB signaling pathways under the concentra-tion of 2.5～200 μg/ml.Conclusion:The ultrasound-assisted method can efficiently isolate CSPFPs with immunomodulatory activity,which provides basic data for the development and application of CSPFPs as an immunostimulant.

Objective:To investigate the role of microRNA-25-5p (miR-25-5p) in melanogenesis, and to explore its underlying mechanisms.Methods:Target genes of miR-25-5p were predicted using the TargetScan database. The interaction between miR-25-5p and the 3' untranslated region (3' UTR) of the RAB11B gene (a member of RAS oncogene family) was validated through a dual-luciferase reporter assay. Post-inflammatory hyperpigmentation (PIH) models were established in female C57BL/6J mice (6 - 8 weeks old) and female brown guinea pigs (4 - 6 weeks old) through daily broadband ultraviolet B (UVB) irradiation on the dorsal skin of the mouse ear or shaved dorsal skin of guinea pigs, while untreated mice and untreated dorsal skin areas of guinea pigs served as control groups. During modeling, these experimental animals received intradermal injections of a miR-25-5p agomir or a miR control agomir. Changes in skin pigmentation were observed, and skin tissue samples were harvested for further analysis after modeling. Melanin content in skin tissues was evaluated using Masson-Fontana staining. Expression of RAB11B and tyrosinase (TYR) in skin tissues was determined using immunohistochemical staining and quantitative real-time PCR (qPCR). Primary human melanocytes were isolated from discarded normal foreskin tissues of healthy males after circumcision. Both primary human melanocytes and human MNT1 melanoma cells were transfected with miR-25-5p mimics or miR control mimics. Relative expression levels of miR-25-5p and RAB11B mRNA were quantified by qPCR using the 2 -ΔΔCt calculation method. In MNT1 cells, miR-25-5p and RAB11B were co-overexpressed to assess their effect on the mRNA expression of RAB11B and TYR. Statistical analysis was conducted using t test or one-way analysis of variance followed by Tukey's post hoc test for multiple comparisons. Results:The bioinformatic prediction and dual-luciferase reporter assay confirmed a binding site for miR-25-5p in the 3′ UTR of the RAB11B gene. In both animal models, the treatment with the miR-25-5p agomir significantly reduced local skin pigmentation compared to the control groups; Masson-Fontana staining showed a marked decrease in the density of melanin granules in the epidermis and dermis in the miR-25-5p agomir groups compared with the miR control agomir groups (mice: 0.050 ± 0.005 vs. 0.087 ± 0.008; guinea pigs: 0.067 ± 0.015 vs. 0.110 ± 0.013; both P < 0.05). Immunohistochemical staining revealed significantly lower expression of RAB11B in mouse skin tissues in the miR-25-5p agomir group than in those in the miR control agomir group (both P < 0.05). qPCR revealed significantly lower mRNA expression of RAB11B and TYR in skin tissues of guinea pigs in the miR-25-5p agomir group than in those in the miR control agomir group (both P < 0.05). Similarly, RAB11B mRNA expression significantly decreased in the miR-25-5p mimics group compared with the miR control mimics group in primary human melanocytes and MNT1 cells (both P < 0.05). In human MNT1 melanoma cells, miR-25-5p overexpression could suppress TYR mRNA expression, whereas co-overexpression of miR-25-5p and RAB11B could reverse this suppression. Conclusion:Overexpression of miR-25-5p could alleviate UVB-induced post-inflammatory hyperpigmentation and inhibit melanogenesis, likely by targeted suppression of RAB11B expression.

Objective:To investigate the role of microRNA-25-5p (miR-25-5p) in melanogenesis, and to explore its underlying mechanisms.Methods:Target genes of miR-25-5p were predicted using the TargetScan database. The interaction between miR-25-5p and the 3' untranslated region (3' UTR) of the RAB11B gene (a member of RAS oncogene family) was validated through a dual-luciferase reporter assay. Post-inflammatory hyperpigmentation (PIH) models were established in female C57BL/6J mice (6 - 8 weeks old) and female brown guinea pigs (4 - 6 weeks old) through daily broadband ultraviolet B (UVB) irradiation on the dorsal skin of the mouse ear or shaved dorsal skin of guinea pigs, while untreated mice and untreated dorsal skin areas of guinea pigs served as control groups. During modeling, these experimental animals received intradermal injections of a miR-25-5p agomir or a miR control agomir. Changes in skin pigmentation were observed, and skin tissue samples were harvested for further analysis after modeling. Melanin content in skin tissues was evaluated using Masson-Fontana staining. Expression of RAB11B and tyrosinase (TYR) in skin tissues was determined using immunohistochemical staining and quantitative real-time PCR (qPCR). Primary human melanocytes were isolated from discarded normal foreskin tissues of healthy males after circumcision. Both primary human melanocytes and human MNT1 melanoma cells were transfected with miR-25-5p mimics or miR control mimics. Relative expression levels of miR-25-5p and RAB11B mRNA were quantified by qPCR using the 2 -ΔΔCt calculation method. In MNT1 cells, miR-25-5p and RAB11B were co-overexpressed to assess their effect on the mRNA expression of RAB11B and TYR. Statistical analysis was conducted using t test or one-way analysis of variance followed by Tukey's post hoc test for multiple comparisons. Results:The bioinformatic prediction and dual-luciferase reporter assay confirmed a binding site for miR-25-5p in the 3′ UTR of the RAB11B gene. In both animal models, the treatment with the miR-25-5p agomir significantly reduced local skin pigmentation compared to the control groups; Masson-Fontana staining showed a marked decrease in the density of melanin granules in the epidermis and dermis in the miR-25-5p agomir groups compared with the miR control agomir groups (mice: 0.050 ± 0.005 vs. 0.087 ± 0.008; guinea pigs: 0.067 ± 0.015 vs. 0.110 ± 0.013; both P < 0.05). Immunohistochemical staining revealed significantly lower expression of RAB11B in mouse skin tissues in the miR-25-5p agomir group than in those in the miR control agomir group (both P < 0.05). qPCR revealed significantly lower mRNA expression of RAB11B and TYR in skin tissues of guinea pigs in the miR-25-5p agomir group than in those in the miR control agomir group (both P < 0.05). Similarly, RAB11B mRNA expression significantly decreased in the miR-25-5p mimics group compared with the miR control mimics group in primary human melanocytes and MNT1 cells (both P < 0.05). In human MNT1 melanoma cells, miR-25-5p overexpression could suppress TYR mRNA expression, whereas co-overexpression of miR-25-5p and RAB11B could reverse this suppression. Conclusion:Overexpression of miR-25-5p could alleviate UVB-induced post-inflammatory hyperpigmentation and inhibit melanogenesis, likely by targeted suppression of RAB11B expression.

Objective:To separate and purify the polysaccharides from Polygonatum filipes,characterize their primary structure and investigate the immunomodulatory effects on RAW264.7 macrophages.Methods:Crude polysaccharides from Polygonatum filipes were extracted by ultrasound assisted method,then Polygonatum filipes polysaccharides(CSPFPs)were obtained after elimination of the proteins with combined papain-Sevag method.The total sugar content was determined by phenol-sulfuric acid method.Structures of CSPFPs were analyzed by fourier transform infrared spectroscopy(FT-IR),high performance gel permeation chromatography(HPGPC)and high performance liquid chromatography(HPLC).Effects of CSPFPs on cell viability,pinocytic activity,TNF-α secretion,MAPK and NF-κB signaling pathways of RAW264.7 cells were explored by MTT,Neutral red,ELISA and Western blot,respectively.Results:Extraction rate of CSPFPs by ultrasound-assisted method was 41.61%,which contained total sugar content of 94.00%.CSPFPs with Mw of 3 125 Da was composed of arabinose(1.85%),galactose(6.14%),glucose(56.41%)and mannose(35.60%).The in vitro experiments showed that CSPFPs were non-cytotoxic and enhanced the pinocytic activity,TNF-α secretion and phosphorylation levels of p38,ERK,JNK,p65,IκB and IKK,indicating the activation of MAPK and NF-κB signaling pathways under the concentra-tion of 2.5～200 μg/ml.Conclusion:The ultrasound-assisted method can efficiently isolate CSPFPs with immunomodulatory activity,which provides basic data for the development and application of CSPFPs as an immunostimulant.

Type 2 diabetes mellitus(T2DM)is a metabolic disorder caused by excessive energy intake and insufficient consumption,which is mainly characterized by the dysfunction of islet β-cell dysfunction and insulin resistance(insulin resistance,IR).In modern medicine,oral administration of T2DM may be accompanied by adverse reactions such as liver and kidney function injury.Studies have proved that traditional Chinese medicine has the significant superiority of multi-target,multi-component and multi-pathway.Mitochondria are the main organelles for cells to provide energy.They supply energy to ATP through oxidative phosphorylation(OXPHOS)reaction to maintain the functional balance of cells.There are closely related to the occurrence of T2DM between mitochondrial dysfunction,abnormal mitochondrial dynamics and impaired mitochondrial autophagy.Mitochondria has been gradually confirmed as the target of traditional Chinese medicine in the treatment of diabetes,but there is a lack of an overview of the effects of traditional Chinese medicine on T2DM by regulating mitochondrial-mediated inflammation.Therefore,this paper reviews the relationship between mitochondrial-mediated inflammation and T2DM,and the mechanism of traditional Chinese medicine to interfere with T2DM by regulating mitochondrial-mediated inflammation.

Specific tumor-targeted gene delivery remains an unsolved therapeutic issue due to aberrant vascularization in tumor microenvironment (TME). Some bacteria exhibit spontaneous chemotaxis toward the anaerobic and immune-suppressive TME, which makes them ideal natural vehicles for cancer gene therapy. Here, we conjugated ZIF-8 metal-organic frameworks encapsulating eukaryotic murine interleukin 2 (Il2) expression plasmid onto the surface of VNP20009, an attenuated Salmonella typhimurium strain with well-documented anti-cancer activity, and constructed a TME-targeted Il2 delivery system named Il2/ZIF-8@Salmonella. Both in vitro and in vivo experiments demonstrated that Il2/ZIF-8@Salmonella maintained the tumor-targeting feature of bacteria, and could be effectively phagocytosed by intratumoral macrophages, thus leading to the expression and secretion of IL2 in TME. The detailed analysis of tumor immune microenvironment (TIME) showed that one dose of combinatorial Il2/ZIF-8@Salmonella achieved synergistic actions on a potent remodeling of TIME, marked by the activation of cytotoxic T cells and M1-polarization of macrophages in TME, thus leading to significant anti-tumor effects in melanoma, orthotopic hepatocellular carcinoma, and pulmonary metastasis models. More importantly, Il2/ZIF-8@Salmonella exhibited high safety to major organs and hematopoietic systems. Taken together, we report a novel plasmid/ZIF-8@Salmonella system that simultaneously achieves effective TME-targeted delivery of therapeutic gene, as well as synergistic re-activation of TIME.

Objective:To investigate the predictive value of plasma atherosclerosis index (AIP) on metabolic associated fatty liver disease (MAFLD) in physical examination population.Methods:It was a cross-sectional study. Total of 97 076 people who completed physical examination in the Health Management Center of the First Affiliated Hospital of Anhui Medical University from January to December 2021 and met the integrity of the study were selected as study subjects. Of the subjects, 31 176 people who met the diagnostic criteria of MAFLD were set as the MAFLD group, and the other 65 900 people were set as the non-MAFLD group. Laboratory indexes, height, weight, blood pressure, liver ultrasound and other indicators in the two groups were collected, and the AIP was calculated. The t-test was used for measurement data and chi-square test was used for counting data to compare the differences between the two groups. Univariate and multivariate logistic regression were used to analyze the influencing factors of MAFLD. The two groups were grouped further according to gender and age, and the difference of AIP prediction efficiency in different groups was evaluated by receiver operating characteristic (ROC) curve. Results:The average age (47.3 years, Z=-31.734), male proportion (76.9%, χ2=7 837.54) and the average value of AIP (0.23, Z=-155.089) in MAFLD group were all higher than those in non-MAFLD group (all P<0.001). After stratified by age, gender, body mass index (BMI), hypertensive or not, hemoglobin A1c (HbA 1c), triglyceride (TG), fasting blood glucose (FBG), the difference of AIP between the two groups was still statistically significant (all P<0.001). Multifactorial regression analysis showed that alanine aminotransferase (ALT) ( OR=1.024), aspartate aminotransferase (AST) ( OR=0.974), serum creatinine (sCr) ( OR=0.975), serum uric acid ( OR=1.004), HbA 1c ( OR=1.231), hemoglobin (HB) ( OR=1.011), platelet(PLT) ( OR=1.002), FBG ( OR=1.131), BMI ( OR=1.419), AIP ( OR=11.318), systolic blood pressure ( OR=1.002), and diastolic blood pressure ( OR=1.012) were independent risk factors for MAFLD (all P<0.001). In the overall population, AIP had an area under the ROC curve (AUC) of 0.808, a cut-off value of 2.045, a sensitivity of 74.5% and a specificity of 72.4%; in the gender subgroup, the AUC was greater in women than in men (0.815 vs 0.764), and the cut-off values, sensitivities and specificities in the two groups were -0.044 vs 0.091, 75.6% vs 72.2%, 73.3% vs 67.6%, respectively; in the age sub-group, the largest AUC (0.848), cut-off value (0.034), sensitivity (79.1%) and specificity (75.3%) were found in the 18-44 years group; the differences were statistically significantin the ROC curve analysis of each group ( P<0.001). Conclusion:AIP is an independent risk factor for MAFLD, which has good predictive value for the occurrence of the disease, and has better predictive effect in women and young groups (18-44 years old).

Objective:To analyze the related factors affecting the inter-arm systolic blood pressure difference (IASBPD) in a physical examination population.Methods:A total of 3 600 adults who underwent physical examination and completed the arteriosclerosis test in the first affiliated hospital of Anhui medical university from January 2019 to June 2021 were selected as the participants by systematic sampling method. Data on age, sex, and history of smoking, heavy drinking, hypertension, type 2 diabetes, and coronary heart disease were recorded. The height, weight, waist circumference, hip circumference, total muscle, total fat and body fat ratio were measured, and body mass index was calculated. The blood pressure of the limbs, ankle brachial index (ABI) were measured synchronously with the arteriosclerosis tester of the Chinese Academy of Sciences, and the IASBPD were calculated. According to the IASBPD value, the participants were divided into two groups: IASBPD<10 mmHg (1 mmHg=0.133 kPa) group and IASBPD≥10 mmHg group, The differences between the two groups were compared, and the related influencing factors of IASBPD were analyzed by multivariate logistic regression.Results:Weight, body mass index, waist circumference, hip circumference, systolic blood pressure, diastolic blood pressure, total muscle, total fat, body fat rate, history of hypertension, proportion of type 2 diabetes mellitus, and proportion of history of coronary heart disease in the IASBPD≥10 mmHg group was higher than that of IASBPD<10 mmHg group [(69.1±11.2) vs (65.3±10.8) kg, (25.6±3.4) vs (24.4±3.3) kg/m 2, (91.3±11.3) vs (87.8±10.6) cm, (98.5±10.4) vs (96.5±9.8) cm, (139.7±20.0) vs (129.7±17.6) mmHg, (80.3±11.6) vs (76.7±10.1) mmHg, (47.5±9.1) vs (45.3±8.8) kg, (19.4±7.0) vs (17.6±6.4) kg, (27.9%±8.5%) vs (26.8%±8.1%), 41.1% vs 29.3%, 16.6% vs 11.7%, 13.1% vs 7.3%] (all P<0.05); ABI was lower than that in IASBPD<10 mmHg group [(1.15±0.11) vs (1.20±0.09)] ( P<0.001). There were no significant differences in height, smoking history and heavy drinking history between the two groups (all P>0.05). Multivariate logistic regression analysis showed that age, systolic blood pressure, body weight and ABI were independent influencing factors of IASBPD≥10 mmHg. Age, systolic blood pressure and body weight were positively correlated with IASBPD≥10 mmHg, while ABI was negatively correlated with IASBPD≥10 mmHg. Conclusion:Increases in age, systolic blood pressure, and body weight and a decrease in ABI are important influencing factors leading to the elevation of IASBPD.

Epithelioid Hemangioendotheliomais a rare, low-grade malignant vascular tumour. It’scalled hepatic epithelioid hemangioendothelioma(HEHE), when it occurs in liver. It can be metastatic and postoperative recurrence. There are few cases have been reported in the literature at home and abroad because of its rarity. The treatment of HEHE is also controversial. With the continuous improvement of surgical techniques of liver transplantation, it is increasingly applied to treat liver failure patients caused by HEHE. Our paper reviews the literature on disease characteristics of HEHE, and liver transplantation for HEHE indications, immunotherapy and prognosis, to illustrate the status and progress of liver transplantation for HEHE.