Objective: To investigate the objective characteristics of tongue manifestation in different stages of damp-heat syndrome in diabetic kidney disease (DKD). Methods: A cross-sectional study enrolled 134 patients with DKD G3-5 stages who met the diagnostic criteria for damp-heat syndrome in DKD. The patients were treated at Dongzhimen Hospital, Beijing University of Chinese Medicine, from May 2023 to January 2024. The patients were divided into three groups: DKD G3, DKD G4, and DKD G5 stage, with 53, 33, and 48 patients in each group, respectively. Clinical general data (gender, age, and body mass index) and damp-heat syndrome scores were collected from the patients. The YZAI-02 traditional Chinese medicine (TCM) AI Tongue Image Acquisition Device was used to capture tongue images from these patients. The accompanying AI Open Platform for TCM Tongue Diagnosis of the device was used to analyze and extract tongue manifestation features, including objective data on tongue color, tongue quality, coating color, and coating texture. Clinical data and objective tongue manifestation characteristics were compared among patients with DKD G3-5 based on their DKD damp-heat syndrome status. Results: No statistically significant difference in gender or body mass index was observed among the three patient groups. The DKD G3 stage group had the highest age (P<0.05). The DKD G3 stage group had a lower score for symptoms of poor appetite and anorexia(P<0.05) than the DKD G5 group. No statistically significant difference was observed in damp-heat syndrome scores among the three groups. Compared with the DKD G5 stage group, the DKD G3 stage group showed a decreased proportion of pale color at the tip and edges of the tongue (P<0.05). The DKD G4 stage group exhibited an increased proportion of crimson at the root of the tongue, a decreased proportion of thick white tongue coating at the root, a decreased proportion of pale color at the tip and edges of the tongue, an increased hue value (indicating color tone) of the tongue color in the middle, an increased brightness value (indicating color lightness) of the tongue coating color in the middle, and an increased thickness of the tongue coating (P<0.05). No statistically significant difference was observed in other tongue color proportions, color chroma values, body characteristics, coating color proportions, coating color chroma values, and coating texture characteristics among the three groups. Conclusion Tongue features differ in different stages of DKD damp-heat syndrome in multiple dimensions, enabling the inference that during the DKD G5 stage, the degree of qi and blood deficiency in the kidneys, heart, lungs, liver, gallbladder, spleen, and stomach is prominent. Dampness is more likely to accumulate in the lower jiao, particularly in the kidneys, whereas heat evil in the spleen and stomach is the most severe. These insights provide novel ideas for the clinical treatment of DKD.

Acute ischemic stroke(AIS)is associated with a high mortality and disability rate.Endovascular therapy(EVT)has emerged as a primary treatment method for achieving vascular reperfusion in large vessel occlusion AIS patients.The cerebral hemodynamic status of patients undergoing reperfusion therapy is closely linked to the extent of cerebral vascular reperfusion and improvement in the ischemic penumbra at the site of injury.Transcranial Doppler(TCD)ultrasound offers non-invasive,reliable,and convenient advantages for evaluating intracranial vessel occlusion or stenosis,guiding treatment decisions,and predicting patient outcomes.The authors reviewed the application progress of TCD in AIS patients.

Objective To investigate the effects of low-to-moderate intensity treadmill exercise on pain and anxiety-like behaviors in rats with chronic constriction injury of the sciatic nerve(CCI),and to explore the neural mechanism of the exercise-related brain-derived neurotrophic factor(BDNF)/tropomyosin receptor kinase B(TrkB)-cAMP-response element binding protein(CREB)pathway in relieving pain and anxiety behaviors in CCI rats.Methods Thirty-two D rats were divided randomly into four groups:sham group,CCI group,sham+exercise(Sham+Exe)group,and CCI+exercise(CCI+Exe)group.Rats in the exercise groups underwent treadmill training for 4 weeks.The paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)were measured before and at different time points after the operation.The elevated plus maze(EPM)and open field test(OFT)were used to evaluate anxiety-like behaviors in the rats.mRNA and protein expression levels of BDNF,TrkB,and CREB in the hippocampus were detected by real-time quantitative reverse transcription PCR and Western Blot,respectively.Results(1)The PWT and PWL on the operative side of the rats were significantly lower in the CCI compared with the sham group at 7,14,21,28,and 35 days after the operation(P＜0.001).The PWT on the ipsilateral side was significantly increased in the CCI+Exe group after 21 days compared with the CCI group(P＜0.05),and the PWL on the ipsilateral side increased significantly after 14 days(P＜0.05).(2)The EPM result showed that rats in the CCI group spent a significantly lower proportion of time in the open arms(P＜0.001)and significantly more time in the closed arms compared with the sham group(P＜0.01).Rats in the CCI+Exe group spent significantly more time in the open arms than the CCI group(P＜0.05).(3)The OFT result showed that rats in the CCI group spent a significantly lower proportion of time in the central area of the open field compared with the sham group(P＜0.001),while the percentage of time was significantly increased in the CCI+Exe group compared with the CCI group(P＜0.05).(4)BDNF,TrkB,and CREB mRNA and protein levels in the hippocampus were significantly lower in the CCI group compared with the sham group(P＜0.05,P＜0.01).Four-week treadmill exercise increased the mRNA and protein expression levels of BDNF,TrkB,and CREB in the hippocampus of CCI rats(P＜0.05).Conclusions Four weeks of treadmill exercise alleviates mechanical and thermal hyperalgesia and anxiety induced by chronic pain in CCI rats.Up-regulation of the BDNF/TrkB-CREB pathway may be one of the mechanisms by which exercise relieves chronic pain and improves anxiety.

To develop a rapid differential detection method for porcine epidemic diarrhea virus(PEDV)and transmissible gastroenteritis virus(PEDV),M gene sequences of PEDV and TGEV were compared,the inner and outer primer pairs and probes were designed according to the highly conserved region.PEDV-Probe was labeled with FAM at5'end and BHQ1 at 3'end.TGEV-Probe was labeled with CY5.5 at the 5'end and BHQ2 at the 3'end.After optimizing the reaction condi-tions and system,a dual fluorescent RT-LAMP assay for PEDV and TGEV rapid identification was established.The amplification could be completed within 60 min in a 63 ℃ water bath and then stopped at 85 ℃ for 10 min.Then the tubes were placed in a multicolor imaging system,and the re-sults could be observed under 520 nm and 690 nm dual channels.There was no cross-reaction when other common swine viral pathogens were detected by this method.The sensitivity of the assay was evaluated with a 10-fold diluted recombinant plasmid as templates.The lowest detection limit was 102 copies/μL recombinant plasmid,which was 10 times more sensitive than the conventional RT-PCR method.Seventy-two PEDV-positive samples,49 TGEV-positive samples,and 40 PEDV and TGEV co-infected samples were detected from 175 anal swab samples of diarrheic piglets by the established method,which were all higher than the detection rates of the conventional RT-PCR method.The dual fluorescent RT-LAMP method established in this study can be used to amplify the target gene in an ordinary water bath without gel electrophoresis,which provides technical sup-port for rapid and convenient differential diagnosis of PED and TGE and simultaneous detection of PEDV and TGEV co-infection.

Objective To investigate the protective effect of polydatin(PLD)on primary cultured rat cortical neurons induced by β-amyloid protein(Aβ25-35)and its possible mechanism.Methods Cortical neurons were isolated from SD rats and primarily cultured,and then divided into control group,PLD group,Aβ25-35 group and Aβ25-35+PLD group.Cell viability was detected by MTT as-say,intracellular and mitochondrial reactive oxygen species(ROS)levels were detected by dichlo-rofluorescein diacetic acid probe or MitoSOX Red staining,opening of mitochondrial membrane permeability transition pore(MPTP)was detected by MPTP kit,and contents of cytochrome C(Cyt C)and mitochondrial transcription factor A(TFAM)were detected by Western blotting.In addition,the activity of intracellular electron transport chain complexes(including complexes Ⅰ,Ⅱ,Ⅲ and Ⅳ)and the level of adenylate triphosphate(ATP)were determined.High performance liquid chromatography(HPLC)was conducted to determine the content of 8-hydroxydeox-yguanosine(8-OHdG)in the mitochondria.Results Compared with the control group,cell vitali-ty,mitochondrial fluorescence intensity,activities of mitochondrial respiratory chain enzymes(complex Ⅰ,Ⅱ,Ⅲ and Ⅳ),intracellular ATP and mitochondrial TFAM expression were signifi-cantly decreased in the cortical neuron from the Aβ25～35 group(P＜0.05,P＜0.01).Compared with the Aβ25-35 group,mitochondrial fluorescence intensity,mitochondrial respiratory chain enzyme activity(complex Ⅰ,Ⅱ,Ⅲ,Ⅳ),intracellular ATP and mitochondrial TFAM expression were obviously increased in the PLD group,and intracellular and mitochondrial ROS levels were nota-bly decreased after cortical neuron exposure for 3,6,12 and 24 h(P＜0.05,P＜0.01).The ratio of cytoplasmic/mitochondrial Cyt C and mitochondrial level of 8-OHdG were statistically lower in the Aβ25-35+PLD group than the Aβ25-35 group[3.02±0.28 vs 5.73±0.45,P＜0.05;(8.07±1.45)× 106 dG vs(16.07±2.29)X 106 dG,P＜0.05].Conclusion PLD can effectively protect cortical neu-rons against Aβ25-35-induced injury,which may be partially by its inhibiting mitochondrial oxida-tive stress and improving mitochondrial function.

Objective:To investigate the risk factors affecting the prognosis of penile cancer after surgery.Methods:We retrospectively analyzed the clinical data on 112 cases of penile cancer treated in Weifang People's Hospital from January 2013 to De-cember 2023.Using the Kaplan-Meier survival curve,χ2 test,Fisher's exact test,and univariate and multivariate Cox risk regression analyses,we compared the clinical characteristics among different groups,and determined the independent prognostic risk factors for cancer-specific survival(CSS)of the patients.Results:The 1-,3-and 5-year CSS rates of the penile cancer patients were 78.2%,66.1% and 63.7%,respectively.Kaplan-Meier analysis indicated a significant correlation of a higher neutrophil-to-lymphocyte ratio(NLR)with a lower CSS rate(P<0.001).Multivariate Cox regression analysis showed high NLR(HR=2.6;95% CI:1.031-6.558;P=0.043)to be an independent risk factor for CSS.Conclusion:Preoperative NLR is an independent risk factor for the prognosis of penile cancer.In addition,older age,farmer or worker occupation,lower education,preoperative lymphocyte-to-monocyte ratio(LMR)≤2.81,preoperative fibrinogen(FIB)≥3.41 g/L,advanced tumor stage and tumor differentiation are associated with the poor prognosis the malignancy.

Objective:To investigate the role of speckle-type POZ(pox virus and zinc finger protein) protein (SPOP) in enterovirus 71 (EV71) infection.Methods:Immunoprecipitation analysis was employed to examine the impact of SPOP on the ubiquitin level of EV71 non-structural protein 2A protease (2A pro), while the phosphorylation level of IFR3 protein was assessed through Western blot. Cells were either overexpressed or knockdown of SPOP, followed by infection with EV71. RT-qPCR was utilized to analyze the transcription level of IFN-β, and the transcription level and protein level of EV71 structural protein VP1 were determined using RT-qPCR and Western blot, respectively. Results:The inhibition of EV71 infection in RD cells was observed following transfection with HA-SPOP. Additionally, it was found that the ubiquitin level of EV71-2A pro increased in a gradient-dependent manner. Subsequent transfection with shSPOP plasmid for endogenous SPOP knockdown resulted in a dose-dependent decrease in the levels of melanoma differentiation-associated gene 5 (MDA5), mitochondrial antiviral signaling (MAVS), and p-IRF3. Conversely, transfection with HA-SPOP plasmid led to a dose-dependent increase in the levels of MDA5, MAVS, and p-IRF3. The expression of SPOP, whether high or low, had an impact on the expression of IFN-β in cells. Additionally, the levels of VP1 mRNA or protein were found to be inhibited or increased. Conclusions:SPOP plays a role in increasing the ubiquitination level of EV71-2A pro, which in turn promotes the phosphorylation level of IRF3 and secretion of IFN-β. This effect is achieved by inhibiting the cleavage of 2A pro against key molecules MAVS and MDA5 in the RLR signaling pathway, ultimately leading to the inhibition of EV71 replication.

ObjectiveTo preliminarily confirm the effective anti-lung cancer sites of Momordicae Semen and Epimedii Folium and study their mechanism of action. MethodOn the basis of preliminary research， the extraction method of Momordicae Semen and Epimedii Folium was optimized and the effective parts were screened under the guidance of pharmacological effects. Different ethanol elution and water elution sites of Momordicae Semen and Epimedii Folium were obtained through adsorption and elution with D101 macroporous resin. The methylthiazolyldiphenyl-tetrazolium bromide （MTT） colorimetric assay was used to detect the effects of total drug extracts and different elution sites on the proliferation of various tumor cell lines， and to screen for the optimal elution site and tumor sensitive strains. Flow cytometry was used to detect the effect of the elution sites of Momordicae Semen and Epimedii Folium on intracellular reactive oxygen species （ROS） and apoptosis in A549 cells. Western blot was used to compare the expressions of tumor protein 53 （p53）， Bcl-2-associated X protein （Bax）， cysteinyl aspartate specific proteinase-3 and 9 （Caspase-3 and Caspase-9） proteins in A549 cells. ResultThe inhibitory effect of Momordicae Semen on the proliferation of A549 cells was better than the kernel of Momordicae Semen， with 50% inhibitory concentration （IC₅₀） being （86.83±2.88） mg·L^-1 and （95.10±18.13） mg·L^-1， respectively. The effect of total extracts of Epimedii Folium on A549 anti proliferation IC₅₀ value was （4.71±0.81） mg·L^-1. The IC₅₀ values of the 40%， 60%， and 80% ethanol and anhydrous ethanol eluted macroporous resins of the total extracts of Momordicae Semen and Epimedii Folium inhibiting A549 proliferation were （45.32±4.38）、 （14.95±0.73）、 （17.07±1.76）、 （14.46±2.35）、 （51.7±2.26）、 （12.37±0.67）、 （20.29±0.93）、 and （3.43±0.91） mg·L^-1， respectively. Compared with the normal group， the 1∶1 combination of Momordicae Semen and Epimedii Folium inhibited A549 cell proliferation in a time-dependent and concentration-dependent manner. Compared with the normal group， 50 mg·L^-1 of the combination of Momordicae Semen and Epimedii Folium significantly increased intracellular ROS expression （P<0.01）. Compared with the normal group， 12.5， 25， 50 mg·L^-1 of the combination of Momordicae Semen and Epimedii Folium significantly increased the expression of A549 cell apoptosis （P<0.01）. Compared with the normal group， 25， 50 mg·L^-1 of the combination of Momordicae Semen and Epimedii Folium significantly increased the expression of p53 in A549 cells （P<0.01）. Compared with the normal group， 12.5， 25， 50 mg·L^-1 of the combination of Momordicae Semen and Epimedii Folium significantly increased the expression of Bax （P<0.01）. Compared with the normal group， 50 mg·L^-1 of the combination of Momordicae Semen and Epimedii Folium significantly reduced the expressions of Caspase-3 and Caspase-9 （P<0.01）. ConclusionThe anti-tumor effect of Momordicae Semen is better than that of the kernel of Momordicae Semen. The anti-tumor substances of Momordicae Semen and Epimedii Folium mainly concentrate in the 60% ethanol to anhydrous ethanol elution site. A549 cells are sensitive to the 1∶1 combination of Momordicae Semen and Epimedii Folium， which can effectively inhibit the cell proliferation. The mechanism may be related to increasing the generation of ROS in A549 cells， promoting their apoptosis， increasing the expressions of apoptotic proteins such as p53 and Bax， and reducing the expressions of Caspase-3 and Caspase-9.

Idiopathic pulmonary fibrosis(IPF)is a chronic,progressive interstitial lung disease.IPF incidence is increasing yearly with high mortality and poor prognoses.At present,IPF pathogenesis remains unclear,and its treatments are limited.The experimental model is important to further study IPF pathogenesis and explore effective preventive and therapeutic measures.In recent years,its modeling method have been continuously developed and optimized.This study summarizes the establishment method and research progress of IPF experimental models in recent years to provide ideas and references for preclinical research to select appropriate experimental models.

OBJECTIVE To investigate the mechanism of Cichorium glandulosum in the treatment of liver fibrosis by using network pharmacology and experimental validation. METHODS A "component-target-pathway" network was constructed with the help of TCMSP, Pubchem, SwissTargetPrediction and Genecards databases, and the STRING database was used to predict the targets of Cichorium glandulosum against liver fibrosis. KEGG and GO enrichment analysis was performed in the DAVID database, and molecular docking of active ingredients and key targets was docked in AUTODOCK. PDGF-BB was used to induce activation of cells and verify the effects of six compounds, including quercetin, quercetin, chicoric acid, caffeic acid, chlorogenic acid, and isochlorogenic acid, on the proliferation, apoptosis, and liver fibrosis indicators of HSC-T6 cells. Western blotting was used to detect the expression of Ras, ERK1, ERK2, C-fos, and JNK proteins in HSC-T6 cells. RESULTS Network pharmacology screened 239 common targets between the components and liver fibrosis, PPI analysis showed that SRC, STAT3, HSP90AA1 and other targets were key targets, KEGG analysis showed that the pathways affected by Cichorium glandulosum included cancer pathways, metabolic pathways, etc. GO analysis predicted that Cichorium glandulosum mainly affected processes such as signal transduction. The molecular docking results showed that the target that could bind well with the components MAPK1, and the components that could bind well with the target aesculetin, caffeic acid and chlorogenic acid. Compared with the model group, the inhibition effect of the six compounds on PDGF-BB-induced HSC-T6 cell activation was stronger, and all 6 compounds had the effects to reverse the index of liver fibrosis, in which aesculetin had the strongest activity(P<0.01). The expression of Ras, ERK1, ERK2, C-fos, and JNK in HSC-T6 cells decreased after the interventions of 6 compounds. CONCLUSION Each component of Cichorium glandulosum has different anti liver fibrosis effects, which are related to the inhibition of ERK/RAS pathway activation.