Objective:To characterize and compare the polysaccharide components of Panax japonicus with its common counterfeits (Dysosma versipellis, Lycopus lucidus and Dysosma pleiantha); To provide a scientific basis for the quality evaluation of polysaccharides of Panax japonicu.Methods:Crude polysaccharides were extracted using water and subsequently precipitated with ethanol. Three batches of total polysaccharides from Panax japonicus, Dysosma versipellis, Lycopus lucidus and Dysosma pleiantha were prepared using the savage deproteinization method. The molecular weight distribution, functional group characteristics and monosaccharide composition of each batch were analyzed using high performance gel filtration chromatography (HPGFC), fourier-transform infrared spectroscopy (FT-IR) and derivatization of 1-phenyl-3-methyl-5-pyrazolinone with high performance liquid chromatography (PMP-HPLC). Using DEAE column chromatography purification and specific enzymolysis, combined with high performance thin layer chromatography and carbohydrate gel electrophoresis, the saccharide profiles of polysaccharides of Panax japonicus and its counterfeits were analyzed and compared.Results:The molecular weight distribution of total polysaccharides from three batches of Panax japonicus exhibited high similarity, with a concentrated distribution ranging from 2.05×10 4 - 1.87×10 3 Da. However, the molecular weight distribution of total polysaccharides from Dysosma versipellis was scattered in regions 5.08×10 6-6.47×10 5 Da and 6.47×10 5-2.05×10 4 Da, while Lycopus lucidus and Dysosma pleiantha was scattered in regions 6.47×10 5-2.05×10 4 Da and 2.05×10 4-1.87×10 3 Da; the infrared spectra of all samples exhibited similarity, indicating that the sugar chains of each polysaccharide were predominantly linked by α-glycosidic bonds, with no significant differences was observed. In terms of monosaccharide composition, the polysaccharides from Panax japonicus, Dysosma versipellis and Dysosma pleiantha were mainly composed of glucose, galactose, arabinose, galacturonic acid, rhamnose and mannose. In contrast, the polysaccharides from Lycopus lucidus were distinct, primarily consisting of galactose and glucose; glycosidic linkage analysis revealed that the polysaccharides purified by DEAE column chromatography from Panax japonicus and its counterfeits were resistant to hydrolysis by β- galactosidase, but could be hydrolyzed by α-amylase and pectinase (except for Lycopus lucidus). The oligosaccharides produced by α-amylase hydrolysis of three batches of Panax japonicus polysaccharides were similar, showing clear differences from those of the counterfeits. The results of pectinase hydrolysis were correlated with the content of uronic acids. Conclusions:The total polysaccharides from Panax japonicus, Dysosma versipellis, Lycopus lucidus and Dysosma pleiantha exhibit significant differences in their molecular weight distributions. The monosaccharide composition of Lycopus lucidus polysaccharides is notably distinct, making it easily distinguishable from other species; purification using DEAE column chromatography, combined with HPTLC and polysaccharide analysis using carbohydrate gel electrophoresis (PACE), effectively differentiates the polysaccharides of Panax japonicus from its counterfeits. This approach provides a valuable reference for the quality analysis of polysaccharides in TCM. Additionally, it lays a foundation for the development and utilization of Panax japonicus polysaccharides.

Objective:To analyse the effect of ultrasound-guided percutaneous liver biopsy and the coaxial biopsy needle tract radiofrequency ablation on patients diagnosed with hepatocellular carcinoma who are considered to be at risk of bleeding.Methods:The data of 117 patients with hepatocellular carcinoma who underwent coaxial biopsy needle tract radiofrequency ablation after ultrasound-guided percutaneous liver biopsy in Peking University First Hospital from March 2019 to April 2023 were retrospectively analysed. There were 95 males and 22 females, with the age of (62.0±11.8) years. A comprehensive analysis was conducted on the following variables: the pre-puncture platelet count, the international standardised ratio, anticoagulation therapy, the haemoglobin (Hb) level, the success rate of the liver puncture, the qualified rate of liver puncture specimens, the number of puncture samples, the length of hospital stay, the Hb level after puncture, bleeding within 10 days post-operation, and complications after ablation, including biliary fistula, hemothorax and organ perforation.Results:Among the 117 patients, 60 cases (51.3%) had an international normalized ratio >1.1, 40 cases (34.2%) had thrombocytopenia, that is, <150×10 9/L, and 17 cases (14.5%) received continuous anticoagulation therapy before the operation. It is evident that all 117 patients successfully completed the ultrasound-guided percutaneous liver biopsy, and that all liver biopsy specimens were qualified. The absence of biliary fistula, hemothorax, organ perforation or death in the patients post-ablation was noted. According to the adverse event evaluation criteria, version 5.0, 113 cases (96.6%) were classified as grade 1 and 4 cases (3.4%) were classified as grade 3. The Hb concentration of patients with minor bleeding (grade 1) prior to puncture was (119.7±22.2) g/L, which was significantly higher than the Hb concentration of patients with severe bleeding (grade 3), (76.0±10.4) g/L ( t=3.92, P=0.010). A meticulous examination of the data revealed that there were no statistically significant differences between the two groups with regard to pre-puncture platelet count, pre-puncture international standardised ratio, pre-puncture proportion of receiving anticoagulant drugs, length of hospital stay and number of puncture samples (all P>0.05). Conclusion:For patients with hepatocellular carcinoma who are at risk of bleeding, ultrasound-guided percutaneous liver biopsy followed by coaxial biopsy needle tract radiofrequency ablation can obtain satisfactory liver tissue samples and is relatively safe. There were differences in hemoglobin levels before puncture among patients with different bleeding after puncture.

Objective To explore the mechanism of total saponin from Panax japonicus(TSPJ)in alleviating neuroinflammation in experimental autoimmune encephalomyelitis(EAE)based on the activation of microglia mediated by MAPK signaling pathway.Methods In vivo experiments,mice were divided into normal group,model group and TSPJ low-and high-dosage groups.EAE mouse model was induced by myelin oligodendrocyte glycoprotein polypeptide.Each medication group was gavaged with corresponding drug solution once a day for 28 days.The number of Iba-1-positive cells was assessed using immunofluorescence,the protein expressions of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1β and MAPK signaling pathway related factors were detected by Western blot.In vitro experiments,murine microglia BV2 were divided into normal group,model group and TSPJ low-and high-dosage groups.LPS+IFN-γ was used to induce M1 polarization of BV2 cells,medication groups were given TSPJ intervention,the content of nitric oxide(NO)in cell supernatant,the expression of M1 microglia markers and MAPK signaling pathway related factors were detected,and ERK and JNK signaling pathway inhibitors were further used to clarify the molecular mechanism of TSPJ antagonizing neuroinflammation.Results The results of in vivo experiments showed that compared with the normal group,the cell body and number of microglia in cerebral cortex of the model group mice increased significantly(P<0.01),and the protein expressions of TNF-α,IL-6,IL-1β,p-p38/p38,p-JNK/JNK,p-ERK1/2/ERK1/2 significantly increased(P<0.05,P<0.01);compared with the model group,the number of microglia in cerebral cortex of TSPJ low-and high-dosage groups significantly decreased(P<0.01),the expressions of TNF-α,IL-6,p-p38/p38,p-JNK/JNK,p-ERK1/2/ERK1/2 protein significantly decreased(P<0.05,P<0.01),and the expression of IL-1β protein in TSPJ high-dosage group significantly decreased(P<0.05).The results of in vitro experiments showed that compared with the normal group,the content of NO in cell supernatant of the model group significantly increased(P<0.01),the protein expressions of CD16,CD86,p-p38/p38,p-JNK/JNK,p-ERK1/2/ERK1/2 significantly increased(P<0.01);compared with the model group,the content of NO in cell supernatant of TSPJ low-and high-dosage groups significantly decreased(P<0.01),and the protein expressions of CD16,CD86,p-p38/p38,p-JNK/JNK,p-ERK1/2/ERK1/2 significantly decreased(P<0.05,P<0.01).TSPJ combined with ERK and JNK pathway inhibitors could further inhibit the expressions of TNF-α and IL-6,but there was no significant difference compared with inhibitors alone.Conclusion TSPJ can inhibit the activation of microglia by regulating MAPK signaling pathway,thereby reducing EAE induced central nervous inflammation.

Objective:To analyse the effect of ultrasound-guided percutaneous liver biopsy and the coaxial biopsy needle tract radiofrequency ablation on patients diagnosed with hepatocellular carcinoma who are considered to be at risk of bleeding.Methods:The data of 117 patients with hepatocellular carcinoma who underwent coaxial biopsy needle tract radiofrequency ablation after ultrasound-guided percutaneous liver biopsy in Peking University First Hospital from March 2019 to April 2023 were retrospectively analysed. There were 95 males and 22 females, with the age of (62.0±11.8) years. A comprehensive analysis was conducted on the following variables: the pre-puncture platelet count, the international standardised ratio, anticoagulation therapy, the haemoglobin (Hb) level, the success rate of the liver puncture, the qualified rate of liver puncture specimens, the number of puncture samples, the length of hospital stay, the Hb level after puncture, bleeding within 10 days post-operation, and complications after ablation, including biliary fistula, hemothorax and organ perforation.Results:Among the 117 patients, 60 cases (51.3%) had an international normalized ratio >1.1, 40 cases (34.2%) had thrombocytopenia, that is, <150×10 9/L, and 17 cases (14.5%) received continuous anticoagulation therapy before the operation. It is evident that all 117 patients successfully completed the ultrasound-guided percutaneous liver biopsy, and that all liver biopsy specimens were qualified. The absence of biliary fistula, hemothorax, organ perforation or death in the patients post-ablation was noted. According to the adverse event evaluation criteria, version 5.0, 113 cases (96.6%) were classified as grade 1 and 4 cases (3.4%) were classified as grade 3. The Hb concentration of patients with minor bleeding (grade 1) prior to puncture was (119.7±22.2) g/L, which was significantly higher than the Hb concentration of patients with severe bleeding (grade 3), (76.0±10.4) g/L ( t=3.92, P=0.010). A meticulous examination of the data revealed that there were no statistically significant differences between the two groups with regard to pre-puncture platelet count, pre-puncture international standardised ratio, pre-puncture proportion of receiving anticoagulant drugs, length of hospital stay and number of puncture samples (all P>0.05). Conclusion:For patients with hepatocellular carcinoma who are at risk of bleeding, ultrasound-guided percutaneous liver biopsy followed by coaxial biopsy needle tract radiofrequency ablation can obtain satisfactory liver tissue samples and is relatively safe. There were differences in hemoglobin levels before puncture among patients with different bleeding after puncture.

Objective To explore the mechanism of total saponin from Panax japonicus(TSPJ)in alleviating neuroinflammation in experimental autoimmune encephalomyelitis(EAE)based on the activation of microglia mediated by MAPK signaling pathway.Methods In vivo experiments,mice were divided into normal group,model group and TSPJ low-and high-dosage groups.EAE mouse model was induced by myelin oligodendrocyte glycoprotein polypeptide.Each medication group was gavaged with corresponding drug solution once a day for 28 days.The number of Iba-1-positive cells was assessed using immunofluorescence,the protein expressions of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1β and MAPK signaling pathway related factors were detected by Western blot.In vitro experiments,murine microglia BV2 were divided into normal group,model group and TSPJ low-and high-dosage groups.LPS+IFN-γ was used to induce M1 polarization of BV2 cells,medication groups were given TSPJ intervention,the content of nitric oxide(NO)in cell supernatant,the expression of M1 microglia markers and MAPK signaling pathway related factors were detected,and ERK and JNK signaling pathway inhibitors were further used to clarify the molecular mechanism of TSPJ antagonizing neuroinflammation.Results The results of in vivo experiments showed that compared with the normal group,the cell body and number of microglia in cerebral cortex of the model group mice increased significantly(P<0.01),and the protein expressions of TNF-α,IL-6,IL-1β,p-p38/p38,p-JNK/JNK,p-ERK1/2/ERK1/2 significantly increased(P<0.05,P<0.01);compared with the model group,the number of microglia in cerebral cortex of TSPJ low-and high-dosage groups significantly decreased(P<0.01),the expressions of TNF-α,IL-6,p-p38/p38,p-JNK/JNK,p-ERK1/2/ERK1/2 protein significantly decreased(P<0.05,P<0.01),and the expression of IL-1β protein in TSPJ high-dosage group significantly decreased(P<0.05).The results of in vitro experiments showed that compared with the normal group,the content of NO in cell supernatant of the model group significantly increased(P<0.01),the protein expressions of CD16,CD86,p-p38/p38,p-JNK/JNK,p-ERK1/2/ERK1/2 significantly increased(P<0.01);compared with the model group,the content of NO in cell supernatant of TSPJ low-and high-dosage groups significantly decreased(P<0.01),and the protein expressions of CD16,CD86,p-p38/p38,p-JNK/JNK,p-ERK1/2/ERK1/2 significantly decreased(P<0.05,P<0.01).TSPJ combined with ERK and JNK pathway inhibitors could further inhibit the expressions of TNF-α and IL-6,but there was no significant difference compared with inhibitors alone.Conclusion TSPJ can inhibit the activation of microglia by regulating MAPK signaling pathway,thereby reducing EAE induced central nervous inflammation.

Objective To explore the roles of iron overload in pro-atherogenic activation of foam cells.Methods RAW264.7 and MOVAS cells were stimulated by oxLDL,ferrimine citrate and deferoxamine respectively.Prussian Blue and Oil Red O staining were used to detect iron deposition and foam cell.CCK-8 test,DHE probe,ELISA,RT-qPCR were performed to detect the cell death rate,reactive oxygen species(ROS)generation,lipid peroxidation molecules[glutathione peroxidase(GSH),glutathione peroxidase 4(GPX4),malondialdehyde(MDA)content]and the mRNA level of ATP binding cassette transporter A1(ABCA1),ATP binding cassette transporter G1(ABCG1),inductible nitris oxide synthase(iNOS),arginase-1(Arg-1),α-smooth muscle actin(α-SMA),smooth muscle 22 alpha(SM22a),osteopontin(OPN),Interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α).Results Iron overload could reduced reverse cholesterol transporters(ABCA1 and ABCG1),promote foam cells generation,increased cell death rate,induced the expression of lipid peroxidation molecules(GSH,GPX4,MDA),and promoted pro-inflammatory M1 marker of macrophage and synthetic marker expression of vascular smooth muscle cell(VSMC)and inflammatory cytokines(IL-1β,TNF-α).Conclusion Iron overload promotes the generation of foam cells derived from macrophages and smooth muscle cells and transform them into pro-atherosclerotic phenotype,aggravates cell lipid peroxidation and inflammatory reaction,which contributes to the progress of atherosclerosis.

Objective:To investigate the prognosis and influencing factors of endoscopic surgery for early glottic carcinoma.Methods:In this retrospective study, we applied the Cox proportional hazards regression model and the random survival forest model to analyze the clinical characteristics of 385 patients [362 males, 23 females, age ranging from 33 to 91 years (62.0±9.6)] who visited the Sixth Medical Center of the General Hospital of the People's Liberation Army from January 2009, to December 2022 and diagnosed with early glottic carcinoma, encompassing variables such as age, gender, T stage, surgical approach, pathological typing, etc. The primary evaluation indicators were overall survival(OS) and disease-free survival rates (DFS). The follow-up duration ranged from 30 to 5,557 days (with a median follow-up time of 1,596 days).Results:After a three-year follow-up, the OS rate for the 385 patients was 95.83%, while the DSF rate was 82.98%. The Cox proportional hazards regression analysis revealed age ( HR=2.35, 95% CI: 1.75 to 3.15, P<0.001) and T staging ( HR=1.59, 95% CI: 1.13 to 2.23, P=0.019) as predominant factors affecting the OS and DFS. The random survival forest model identified poor tumor differentiation, and high expression of P53 and Ki-67 as predictors of inferior prognosis. Conclusion:Endoscopic surgery for early glottic carcinoma yields favorable short-term OS and reduces short-term recurrence rates, with T-stage emerging as a pivotal factor influencing recurrence. Tumors with poor differentiation and elevated expression of P53 may be indicative of an increased risk of recurrence.

The widespread of tigecycline resistance gene tet(X4) has a serious impact on the clinical efficacy of tigecycline. The development of effective antibiotic adjuvants to combat the looming tigecycline resistance is needed. The synergistic activity between the natural compound β-thujaplicin and tigecycline in vitro was determined by the checkerboard broth microdilution assay and time-dependent killing curve. The mechanism underlining the synergistic effect between β-thujaplicin and tigecycline against tet(X4)-positive Escherichia coli was investigated by determining cell membrane permeability, bacterial intracellular reactive oxygen species (ROS) content, iron content, and tigecycline content. β-thujaplicin exhibited potentiation effect on tigecycline against tet(X4)-positive E. coli in vitro, and presented no significant hemolysis and cytotoxicity within the range of antibacterial concentrations. Mechanistic studies demonstrated that β-thujaplicin significantly increased the permeability of bacterial cell membranes, chelated bacterial intracellular iron, disrupted the iron homeostasis and significantly increased intracellular ROS level. The synergistic effect of β-thujaplicin and tigecycline was identified to be related to interfere with bacterial iron metabolism and facilitate bacterial cell membrane permeability. Our studies provided theoretical and practical data for the application of combined β-thujaplicin with tigecycline in the treatment of tet(X4)-positive E. coli infection.
Humans ; Tigecycline/pharmacology* ; Escherichia coli/metabolism* ; Reactive Oxygen Species/therapeutic use* ; Plasmids ; Anti-Bacterial Agents/metabolism* ; Escherichia coli Infections/microbiology* ; Bacteria/genetics* ; Microbial Sensitivity Tests

Objective: To systematically evaluate the efficacy and safety of Bifidobacteria in preventing caries. Methods : Databases including PubMed, Embase, The Cochrane Library, Web of Science, Scopus, Clinicaltrials. gov, CNKI, WanFang Data and VIP were electronically searched from inception to April 2020 to collect randomized controlled trials of Bifidobacterium for caries. Meta-analysis was performed using Revman 5.4 software. Results: In total, 10 randomized controlled trials (RCT) of 518 patients, including 262 in the test group and 256 in the control group, were included. Meta-analysis results reveal no statistically significant differences in salivary Streptococcus mutans counts (SMD=-0.31, 95%CI -0.66 to 0.04, P=0.08) (RR=0.53, 95%CI 0.17 to 1.66, P=0.28) and salivary Lactobacilli counts (SMD=-0.07, 95%CI -0.39 to 0.26, P=0.69) (RR=0.87, 95%CI 0.59 to 1.29, P=0.50). No statistical differences in the counts of Streptococcus mutans and Lactobacillus counts were noted in dental plaque, and no statistical difference in the occurrence of caries in deciduous teeth. Three of the 10 RCTS included in this study did not report adverse events, 5 had no adverse reactions, and 2 reported gastrointestinal discomfort. Conclusion Current evidence suggests that Bifidobacteria do not effectively reduce Streptococcus mutans counts and Lactobacillus counts in saliva and dental plaque, or reduce the occurrence of caries in deciduous teeth. The safety of this treatment also requires further investigation.

Objective: To compare the morphology of the crown roots of upper and lower canines in patients with skeletal Class Ⅰ, Ⅱ, and Ⅲ malocclusions by CBCT and to provide guidance for the clinical treatment of orthodontics. Methods: Randomly selected patients with permanent occlusal malocclusion who had undergone CBCT and X-ray skull lateral radiographs were Classified according to the ANB angle size: group Ⅰ, group Ⅱ, and group Ⅲ. Three-dimensional reconstruction was used to obtain the median sagittal section images of the right upper and lower canines. The crown root angle, crown root deflection distance, and lip tangent angle at the center of the clinical crown were used as indicators for measurement and analysis with the use of AutoCAD software. Results : The difference in the crown-root skew distance between different sagittal face types, including upper canines (F=3.335, P=0.042), lower canines (F=3.745, P=0.029) crown root angles and upper canines (F=3.312, P=0.043), and lower canines (F=3.641, P= 0.032), was statistically significant (P < 0.05). The crown root angle of the maxillary canine in group Ⅰ was larger than that in group Ⅱ, and the deflection distance of the crown root was negative and the absolute value was lager in group Ⅰ than in group Ⅱ (P < 0.05). The deflection distance was positive and greater in group Ⅲ than in groups Ⅰ and Ⅱ (P < 0.05). There was no significant difference in the maxillary canine crown-labial tangent angle between the different sagittal facial misalignment groups (P > 0.05). Conclusion Differences in the morphology of canines were found among subjects with skeletal Class Ⅰ, Ⅱ, and Ⅲ malocclusions. The root of the upper canine in Class Ⅰ malocclusions was relatively closer to the labial side of the crown than that in Class Ⅱ malocclusions. The root of the lower canine in Class Ⅲ malocclusions was the closest to the lingual side of the crown among the three Classes.