OBJECTIVE: To observe the effects of Tongdu Tiaoshen acupuncture (for unblocking the obstruction in the governor vessel and regulating the spirit) on depression-like behavior and the hippocampal Endophilin A1/reactive oxygen species (ROS) pathway in the chronic unpredictable mild stress (CUMS) model rats, and explore the mechanism of this therapy for depression. METHODS: Forty-eight male SD rats of SPF grade were randomly divided into a normal group (n=12) and a modeling group (n=36). In the modeling group, CUMS was performed to establish depression model. The successfully-modeled rats were randomized into a model group, a Tongdu Tiaoshen acupuncture group (referred to as the acupuncture group), and a fluoxetine group, with 12 rats in each group. In the acupuncture group, "Baihui" (GV20), "Shenting" (GV24), "Shuigou" (GV26) and "Dazhui" (GV14) were stimulated with acupuncture. This intervention measure was delivered once a day, continuously for 6 days; it was discontinued on day 7 and was completed in 28 days. In the fluoxetine group, intragastric administration was done with fluoxetine solution (2.1 mg/kg), once a day, and for 28 consecutive days. Before and after modeling, and after intervention completion, the body mass, sucrose preference rate and the total distance of movement and the boxes of horizontal crossing in the open field experiment were observed in each group. After intervention, using HE staining, the hippocampal neuron morphology was observed; using Nissl staining, the hippocampal Nissl body number was counted. The hippocampal mitochondria was observed under transmission electron microscopy. The average fluorescence intensity of ROS in hippocampal was determined using flow cytometry. With Western blot method, the protein expression of Endophilin A1, growth associated protein 43 (GAP-43), and brain-derived neurotrophic factor (BDNF) in hippocampal was detected; and with RT-qPCR method, the mRNA expression of Endophilin A1, GAP-43, and BDNF was recorded. Using the immunofluorescence, the average fluorescence intensity of Endophilin A1, GAP-43, and BDNF in hippocampal tissue was determined. RESULTS: Compared with the normal group, in the model group, the body mass, sucrose preference rate, and the total distance of movement and the boxes of horizontal crossing in the open field experiment decreased (P<0.01); the hippocampal neuronal structure was unclear, the matrix was relatively loose, and the number of Nissl body decreased (P<0.01); mitochondrial structure was disarranged, the outer membrane was ruptured, mitochondrial cristae was irregular or missed; the average fluorescence intensity of ROS in hippocampal tissue, the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue increased (P<0.01), while the protein and mRNA expression of GAP-43 and BDNF and its average fluorescence intensity decreased (P<0.01). Compared with the model group, the acupuncture group and the fluoxetine group showed the increase in body mass, sucrose preference rate, the total distance of movement and the boxes of horizontal crossing in the open field experiment (P<0.05, P<0.01); the hippocampal neuronal structure became relatively clear, the matrix was relatively dense, and the number of Nissl body was elevated (P<0.01); mitochondrial structure got normal and disarranged slightly, the average fluorescence intensity of ROS in hippocampal tissue, the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue were reduced (P<0.01), while the protein and mRNA expression of GAP-43 and BDNF and the average fluorescence intensity rose (P<0.01, P<0.05). Compared with the fluoxetine group, the acupuncture group presented the increase in the average fluorescence intensity of ROS, the protein expression and the average fluorescence intensity of Endophilin A1, the protein expression of GAP-43 and the mRNA expression of BDNF (P<0.01, P<0.05), and the decrease of the protein expression and the average fluorescence intensity of BDNF, the mRNA expression of Endophilin A1, and the average fluorescence intensity of GAP-43 (P<0.01, P<0.05). CONCLUSION Tongdu tiaoshen acupuncture alleviates depression-like behaviors in CUMS model rats and protects hippocampal neurons, which may be related to suppressing Endophilin A1/ROS signaling pathway and attenuating oxidative stress reactions.
Animals ; Male ; Hippocampus/metabolism* ; Acupuncture Therapy ; Rats, Sprague-Dawley ; Rats ; Depression/psychology* ; Humans ; Reactive Oxygen Species/metabolism* ; Disease Models, Animal ; Acupuncture Points

Nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) is a cytosolic pattern recognition receptor that recognizes multiple pathogen-associated molecular patterns and damage-associated molecular patterns. It is a cytoplasmic immune factor that responds to cellular stress signals, and it is usually activated after infection or inflammation, forming an NLRP3 inflammasome to protect the body. Aberrant NLRP3 inflammasome activation is reportedly associated with some inflammatory diseases and metabolic diseases. Recently, there have been mounting indications that NLRP3 inflammasomes play an important role in liver injuries caused by a variety of diseases, specifically hepatic ischemia/reperfusion injury, hepatitis, and liver failure. Herein, we summarize new research pertaining to NLRP3 inflammasomes in hepatic injury, hepatitis, and liver failure. The review addresses the potential mechanisms of action of the NLRP3 inflammasome, and its regulation in these liver diseases.
Humans ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Inflammasomes/physiology* ; Animals ; Liver Diseases/metabolism* ; Liver/metabolism* ; Reperfusion Injury/metabolism*

ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design（QbD） and to assess its health risk. MethodsTaking whole iron content， Fe²⁺ dissolution content and looseness as critical quality attributes（CQAs）， and calcination temperature， calcination time， spreading thickness and particle size as critical process parameters（CPPs） determined by the failure mode and effect analysis（FMEA）， the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation（AHP-CRITIC） hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry， and the health risk assessment was carried out by daily exposure（EXP）， target hazard quotient（THQ） and lifetime cancer risk（LCR）， and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃， calcination time of 1 h， particle size of 0.2-0.5 cm， spreading thickness of 1 cm， and vinegar quenching for 1 time［Haematitum-vinegar（10：3）］. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1， while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10^-6-1×10^-4， and the LCR of the rest was<1×10^-6， and the LCRs of calcined Haematitum were all<1×10^-6， indicating that the carcinogenic risk of calcined Haematitum was low， but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg were formulated as 1 014， 25， 17， 27， 7 mg·kg^-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible， and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.

ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design（QbD） and to assess its health risk. MethodsTaking whole iron content， Fe²⁺ dissolution content and looseness as critical quality attributes（CQAs）， and calcination temperature， calcination time， spreading thickness and particle size as critical process parameters（CPPs） determined by the failure mode and effect analysis（FMEA）， the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation（AHP-CRITIC） hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry， and the health risk assessment was carried out by daily exposure（EXP）， target hazard quotient（THQ） and lifetime cancer risk（LCR）， and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃， calcination time of 1 h， particle size of 0.2-0.5 cm， spreading thickness of 1 cm， and vinegar quenching for 1 time［Haematitum-vinegar（10：3）］. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1， while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10^-6-1×10^-4， and the LCR of the rest was<1×10^-6， and the LCRs of calcined Haematitum were all<1×10^-6， indicating that the carcinogenic risk of calcined Haematitum was low， but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg were formulated as 1 014， 25， 17， 27， 7 mg·kg^-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible， and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.

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 identify risk factors for mucus plug formation and to construct a nomogram-based predictive model in children with adenovirus pneumonia.Methods:In this retrospective cohort study, 149 pediatric patients diagnosed with adenovirus pneumonia were included. All patients underwent bronchoscopy at the Department of Respiratory Tianjin Children′s Hospital Machang District, between January and December 2024. Their demographic characteristics, clinical manifestations, laboratory findings, imaging features, and treatment details were analyzed.The information of patients were collected about clinical manifestations, laboratory findings, imaging features, and treatment information. Patients were divided into a mucus plug group and a non-mucus plug group based on bronchoscopic findings. Independent risk factors were determined using multivariable Logistic regression. And a predictive nomogram was subsequently developed. Model performance was evaluated using the area under the receiver operating characteristic curve and the Hosmer-Lemeshow goodness-of-fit test.Results:Of the 149 enrolled patients (84 boys, 65 girls), 49 cases were classified into the mucus plug group and 100 cases into the non-mucus plug group. Compared with their counterparts, children in the mucus plug group were older on admission ( P<0.05), more likely to have a recent history of lower respiratory tract infection, and exhibited a higher prevalence of consolidation or bronchiolitis patterns on chest imaging (all P<0.05), while preoperative corticosteroid use was less frequent ( P<0.05). Multivariable Logistic regression identified age on admission ( OR=1.14, 95% CI 1.01-1.29, P=0.031), lower respiratory tract infection ( OR=5.23, 95% CI 2.15-13.56, P<0.001), radiographic consolidation ( OR=2.64, 95% CI 1.10-6.61, P=0.032), and bronchiolitis pattern ( OR=2.39, 95% CI 1.04-5.60, P=0.042) as independent risk factors, whereas pre-bronchoscopy corticosteroid use was a protective factor ( OR=0.26, 95% CI 0.10-0.63, P=0.004). The nomogram predictive model demonstrated strong discriminative ability (area under the curve=0.83) and good calibration (Hosmer-Lemeshow P=0.358). Conclusions:The independent risk factors for mucus plug formation include older age, a recent history of lower respiratory tract infection, and chest imaging features of consolidation or bronchiolitis in pediatric adenovirus pneumonia. Pre-bronchoscopy corticosteroid therapy is a protective effect. The developed nomogram demonstrates favorable predictive performance and may facilitate early identification and timely intervention.

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.

The receptor-binding region(RBD)of the spike protein(S)on the surface of porcine epi-demic diarrhea virus(PEDV)is a critical structural domain mediating viral invasion of host cells and serves as a key target for inducing neutralizing antibodies.In order to prepare antibodies that can be used to study the biological function of PEDV RBD and develop novel diagnostic and thera-peutic methods,recombinant RBD protein expressed in Sf9 insect cells was utilized as an immuno-gen to immunize BALB/c mice.Monoclonal antibodies(mAbs)were generated via hybridoma tech-nology,and positive hybridoma clones were screened using indirect ELISA.The reactivity of the mAbs was subsequently characterized.The results of ELISA,Western blot,and indirect immuno-fluorescence assay(IFA)showed that three monoclonal antibodies screened(3E5,4F9 and 5A5)had good reactivity with the virus and RBD protein.Antibody subtype results showed that 3E5 and 4F9 were of IgG1 subtypes and 5A5 was of IgM subtype.Neutralization assay further revealed that 3E5 monoclonal antibody had viral neutralizing activity.In this study,three monoclonal antibodies against PEDV RBD proteins were successfully prepared,providing the basis for the study of the bi-ological function of RBD proteins,PEDV serologic detection and vaccine development.

Objective To explore the bronchoscopic manifestations of invasive pulmonary fungal disease(IPFD)and evaluate the safety and efficacy of interventional therapy effect.Methods A retrospective analysis was conducted on the clinical data of 35 patients with IPFD who underwent bronchoscopy from May 12,2018 to May 12,2025 to observe the bronchoscopic manifestations and interventional therapy of IPFD.Results 35 patients' clinical data were collected,including 22 males and 13 females,with an age of(53±14)years.The underlying diseases included:10 cases of hematological malignancies,5 cases of long-term use of glucocorticoids,4 cases of type 2 diabetes,2 cases of lung malignancies,and 1 case of organ transplant patient.Chest CT showed that there were 19 cases of lung lesions involving a single lobe and 16 cases involving multiple lobes.The main manifestations under bronchoscopy were 29 cases with mucosal hyperemia and edema(82.9％),22 cases with necrotic substances blocking the lumen(62.9％),17 cases with a large amount of viscous purulent secretions(48.6％),16 cases with partial bronchial stenosis or occlusion(45.7％),9 cases with mucosal necrosis(25.7％),5 cases with mucosal hemorrhage(14.3％),and 3 cases with fungal globules(8.6％).Among them,32 cases(91.4％)received systemic antifungal treatment,17 cases(48.6％)received local amphotericin B perfusion treatment by bronchoscopy,10 cases(28.6％)had the lesion removed by biopsy forceps,6 cases(17.1％)had the lesion frozen by cryoprobe,and 4 cases(11.4％)received argon plasma coagulation treatment.The clinical symptoms of 28 cases(80.0％)improved significantly,and the lung imaging lesions shrank or regressed.Four cases(11.4％)had stable lesions.Three cases(8.6％)did not complete the treatment.Conclusion IPFD is more common in immunosuppressed hosts,bronchoscopy often presents with necrotic substances blocking the lumen,accompanied by purulent secretions,bronchial lumen stenosis,local mucosal hyperemia,edema or necrosis.Systemic antifungal drugs combined with interventional therapy under bronchoscopy have high safety and good therapeutic effects.

Objective To explore the bronchoscopic manifestations of invasive pulmonary fungal disease(IPFD)and evaluate the safety and efficacy of interventional therapy effect.Methods A retrospective analysis was conducted on the clinical data of 35 patients with IPFD who underwent bronchoscopy from May 12,2018 to May 12,2025 to observe the bronchoscopic manifestations and interventional therapy of IPFD.Results 35 patients' clinical data were collected,including 22 males and 13 females,with an age of(53±14)years.The underlying diseases included:10 cases of hematological malignancies,5 cases of long-term use of glucocorticoids,4 cases of type 2 diabetes,2 cases of lung malignancies,and 1 case of organ transplant patient.Chest CT showed that there were 19 cases of lung lesions involving a single lobe and 16 cases involving multiple lobes.The main manifestations under bronchoscopy were 29 cases with mucosal hyperemia and edema(82.9％),22 cases with necrotic substances blocking the lumen(62.9％),17 cases with a large amount of viscous purulent secretions(48.6％),16 cases with partial bronchial stenosis or occlusion(45.7％),9 cases with mucosal necrosis(25.7％),5 cases with mucosal hemorrhage(14.3％),and 3 cases with fungal globules(8.6％).Among them,32 cases(91.4％)received systemic antifungal treatment,17 cases(48.6％)received local amphotericin B perfusion treatment by bronchoscopy,10 cases(28.6％)had the lesion removed by biopsy forceps,6 cases(17.1％)had the lesion frozen by cryoprobe,and 4 cases(11.4％)received argon plasma coagulation treatment.The clinical symptoms of 28 cases(80.0％)improved significantly,and the lung imaging lesions shrank or regressed.Four cases(11.4％)had stable lesions.Three cases(8.6％)did not complete the treatment.Conclusion IPFD is more common in immunosuppressed hosts,bronchoscopy often presents with necrotic substances blocking the lumen,accompanied by purulent secretions,bronchial lumen stenosis,local mucosal hyperemia,edema or necrosis.Systemic antifungal drugs combined with interventional therapy under bronchoscopy have high safety and good therapeutic effects.