ObjectiveTo investigate the mechanism of action of Kaixinsan in the treatment of Alzheimer's disease （AD） based on network pharmacology， molecular docking， and animal experimental validation. MethodsThe Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） and the Encyclopedia of Traditional Chinese Medicine（ETCM） databases were used to obtain the active ingredients and targets of Kaixinsan. GeneCards， Online Mendelian Inheritance in Man（OMIM）， TTD， PharmGKB， and DrugBank databases were used to obtain the relevant targets of AD. The intersection （common targets） of the active ingredient targets of Kaixinsan and the relevant targets of AD was taken， and the network interaction analysis of the common targets was carried out in the STRING database to construct a protein-protein interaction（PPI） network. The CytoNCA plugin within Cytoscape was used to screen out the core targets， and the Metascape platform was used to perform gene ontology（GO） functional enrichment analysis and Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment analysis. The “drug-active ingredient-target” interaction network was constructed with the help of Cytoscape 3.8.2， and AutoDock Vina was used for molecular docking. Scopolamine （SCOP） was utilized for modeling and injected intraperitoneally once daily. Thirty-two male C57/BL6 mice were randomly divided into blank control （CON） group （0.9% NaCl， n=8）， model （SCOP） group （3 mg·kg^-1·d^-1， n=8）， positive control group （3 mg·kg^-1·d^-1 of SCOP+3 mg·kg^-1·d^-1 of Donepezil， n=8）， and Kaixinsan group （3 mg·kg^-1·d^-1 of SCOP+6.5 g·kg^-1·d^-1 of Kaixinsan， n=8）. Mice in each group were administered with 0.9% NaCl， Kaixinsan， or Donepezil by gavage twice a day for 14 days. Morris water maze experiment was used to observe the learning memory ability of mice. Hematoxylin-eosin （HE） staining method was used to observe the pathological changes in the CA1 area of the mouse hippocampus. Enzyme linked immunosorbent assay（ELISA） was used to determine the serum acetylcholine （ACh） and acetylcholinesterase （AChE） contents of mice. Western blot method was used to detect the protein expression levels of signal transducer and activator of transcription 3（STAT3） and nuclear transcription factor（NF）-κB p65 in the hippocampus of mice. ResultsA total of 73 active ingredients of Kaixinsan were obtained， and 578 potential targets （common targets） of Kaixinsan for the treatment of AD were screened out. Key active ingredients included kaempferol， gijugliflozin， etc.. Potential core targets were STAT3， NF-κB p65， et al. GO functional enrichment analysis obtained 3 124 biological functions， 254 cellular building blocks， and 461 molecular functions. KEGG pathway enrichment obtained 248 pathways， mainly involving cancer-related pathways， TRP pathway， cyclic adenosine monophosphate（cAMP） pathway， and NF-κB pathway. Molecular docking showed that the binding of the key active ingredients to the target targets was more stable. Morris water maze experiment indicated that Kaixinsan could improve the learning memory ability of SCOP-induced mice. HE staining and ELISA results showed that Kaixinsan had an ameliorating effect on central nerve injury in mice. Western blot test indicated that Kaixinsan had a down-regulating effect on the levels of NF-κB p65 phosphorylation and STAT3 phosphorylation in the hippocampal tissue of mice in the SCOP model. ConclusionKaixinsan can improve the cognitive impairment function in SCOP model mice and may reduce hippocampal neuronal damage and thus play a therapeutic role in the treatment of AD by regulating NF-κB p65， STAT3， and other targets involved in the NF-κB signaling pathway.

This study aims to construct a recombinant vaccine containing the tetanus toxin C frag-ment(HC)with a built-in adjuvant of metallothionein 3(MT-3)and evaluate its immune effect in mice.The gene sequences of MT-3 and HC were fused via a linker to create the pET30a-MT-3-HC recombinant plasmid,which was then transformed into E.coli BL21(DE3)competent cells.The re-combinant plasmid was confirmed through double enzyme digestion.The M3C recombinant protein expression was induced,identified by SDS-PAGE and Western blot,and purified using Ni-NTA af-finity chromatography.Five groups of vaccines,including PBS,HC,TT(tetanus toxoid),M3C,and M3C+CpG(composite adjuvant),were administered to mice via intramuscular injection at 7-day intervals for three immunizations.Blood samples were periodically collected from the tail vein.ELISA was used to measure changes in specific IgG antibody titers in the serum,and on day 28,antibody subtypes(IgG1,IgG2a,IgG2b,IgG3,and IgM)and cytokine levels(IL-4,IFN-γ)were also measured.The results demonstrated that the pET30a-MT-3-HC recombinant plasmid was cor-rectly constructed,and the M3C recombinant protein was highly expressed in the supernatant fol-lowing ultrasonic disruption of the induced bacterial culture,with a single band observed post-puri-fication.ELISA results showed that the titer of specific IgG antibodies in the M3C+CpG group peaked at 3.54×105 14 days after the third immunization,which was 141 times,70 times,and 6.6 times higher than that in HC,TT,and TT groups,respectively.Antibody subtype analysis revealed significantly higher specific antibody subtype titers in the M3C and M3C+CpG groups compared to the PBS,HC,and TT groups(P＜0.05),with the M3 C group showing an IgG1/IgG2a ratio greater than 1,and the M3C+CpG group having an IgG1/IgG2a ratio of approximately 1.Serum concentrations of IFN-γ and IL-4 in the M3C and M3C+CpG groups were also significantly higher than those in the other experimental groups(P＜0.05).These results showed that the recombinant antigen containing MT-3 built-in adjuvant and tetanus toxin C fragment was successfully expressed and showed strong immunogenicity,which laid the experimental foundation for the development of this recombinant vaccine.

Objective:To explore the effect of transcranial direct current stimulation (tDCS) on dysphagia in hemispheric stroke patients.Methods:Sixty-two hemispheric stroke patients with dysphagia were randomized into an ipsilateral group, a contralateral group and a bilateral group with 20 in each group. The ipsilateral and contralateral groups received tDCS over their ipsilesional and contralesional hemispheres, respectively, while in the bilateral group it was over both hemispheres. That was followed by conventional swallowing therapy. Before and after 2 weeks of the treatment, swallowing function was assessed using the modified Mann Assessment of Swallowing Ability (MMASA) and a Swallow Severity scale (SSS). Linear regressions were evaluated to highlight the factors most influencing recovery from post-stroke hemispheric dysphagia.Results:After the treatments, the average MMASA and SSS scores had increased significantly in all three groups. There was no significant difference in the average post-treatment MMASA and SSS scores between the ipsilateral and contralateral groups, but the bilateral group showed significantly better average post-treatment MMASA and SSS scores compared to the other two groups. Linear regression analysis confirmed that the tDCS protocol (group allocation) was a significant predictor of recovery.Conclusion:Bilateral tDCS can effectively promote the recovery of swallowing function after a hemispheric stroke. It demonstrates greater therapeutic benefits than unilateral tDCS.

Objective To explore the regulatory effect of Lingzhu Granules on the PI3K/Akt signaling pathway in the rats with polycystic ovary syndrome(PCOS)accompanied by insulin resistance.Methods Fe-male Sprague-Dawley(SD)rats aged 3 weeks d old were selected to establish the model by using the DHEA method.The intervention treatment was administered by different doses of Lingzhu Granules and the homeostasis model as-sessment of insulin resistance(HOMA-IR)was calculated.The relative expression levels of PI3K,Akt,GSK-3β and GLUT4 mRNA in the ovaries was detected by RT-PCR.The expression levels of PI3K and Akt were detected by Western blot.Results Compared with the model control group,the rat body weight in the medium and high doses Lingzhu Granules groups was significantly reduced(P＜0.05).The fasting blood glucose,fasting insulin and HOMA-IR index in the Lingzhu Granules low,medium and high doses groups were significantly lower than those in the model control group,and the differences were statistically significant(P＜0.01).The mRNA relative expression levels of PI3K,Akt,GSK-3β and GLUT4 in the Lingzhu Granules medium and high doses groups all were higher than those in the model control group(P＜0.05).The relative expression levels of PI3K and Akt in the ovarian tissue in the Lingzhu Granules low,medium and high doses groups were higher than those in the model control group,and the differences were statistically significant(P＜0.05).Conclusion Lingzhu Granules could effectively improve the insulin resistance status of PCOS rats by enhancing the synthesis of PI3K and Akt.

Objective:To analyze the clinical phenotypes and genetic characteristics of pediatric patients with neurofibromatosis type 1 (NF1).Methods:A cross-sectional study was adopted. Clinical and imaging data of 25 pediatric patients diagnosed as having NF1 in Department of Pediatric Neurology, Linyi People's Hospital Affiliated to Shandong Second Medical University from January 2024 to July 2025 were collected. Whole exome sequencing and Sanger sequencing were used to conduct genetic testing on the pediatric patients and his/her parents. Protein 3D modeling of the domestic and foreign unreported variations was conducted using SWISS-MODEL software.Results:Among the 25 pediatric patients with NF1, 14 were male (56%) and 11 were female (44%), with age ranging from 8 months to 18 years. All pediatric patients exhibited café-au-lait macules, and 7 (28%) presented with plexiform neurofibromas. Genetic test identified 4 types of NF1 variants: nonsense variant ( n=11, 44%), frameshift variant ( n=9, 36%), missense variant ( n=3, 12%), and splice-site variant ( n=2, 8%). Importantly, 5 novel NF1 variants were discovered, including c.3455T>A, c.3709dupG, c.2665_2684del, c.7092_7095delinsTA, and c.3260del. Three pediatric patients inherited NF1 variant from their parents, while the remaining 22 harbored de novo mutation. Conclusion:NF1 exhibits a broad clinical spectrum, primarily affecting the skin and nervous system; this study identifies 5 previously unreported variants, expanding the genetic profile of NF1.

Objective To investigate the mechanism of action and potential biomarkers of Jiangzhi Qingshen capsules in the treatment of dyslipidaemia based on clinical lipid metabolomics.Methods 30 patients with dyslipidaemia from Zhengzhou Hospital of Chinese Medicine were collected as the test group,and their lipid levels before and after taking Jiang Zhi Qing Shen capsule for 12 weeks were compared.Another 30 healthy patients were enrolled in the physical examination department of Zhengzhou Hospital of Chinese Medicine,and metabolomics investigation was carried out on plasma samples of the test group before and after the treatment as well as those of the healthy patients by LC-MS/MS technology.The differences between the groups were compared by multivariate statistical analysis,and the potential biomarkers were identified by HMDB and lipidblast databases,so as to clarify the possible pathways and targets for the treatment of mild dyslipidaemia by Jiangzhi Qingshen capsules.Results Jiangzhi Qingshen capsules could improve patients' blood lipid level,BMI and abdominal circumference significantly(P<0.05).And metabolomics results showed that 29 lipid metabolites in the plasma of the treated patients were dialled back,which involved cholesterol metabolism,fat digestion and absorption,glycerol-phospholipid metabolism and other biological processes.Conclusion The efficacy of Jiangzhi Qingshen capsules in treating dyslipidaemia was confirmed,and its mechanism of action might be related to the regulation of lipid metabolism.Metabolites such as acylcarnitine(Acar),phosphatidylethanolamine(PE)and phosphatidylcholine(PC)were expected to be used as the biomarkers of dyslipidaemia,so as to provide objective scientific basis for the lipid-lowering capsule,and to facilitate the promotion of its application.

ObjectiveThe study aims to investigate the intervention effect of modified Xiangsha Liujunzitang （M-XSLJZ） on intestinal-derived lipopolysaccharide （LPS）-activated Kupffer cell inflammation in rats with hyperlipidemia spleen deficiency syndrome. MethodsSeventy male SD rats were randomly divided into seven groups （n=10）： blank control （CON）， high-fat diet without spleen deficiency （HFD）， high-fat diet with spleen deficiency （SD-HFD）， M-XSLJZ low-， medium-， and high-dose groups （XS-L， XS-M， XS-H）， and western medicine control （R）. Spleen deficiency was induced in SD-HFD， XS-L， XS-M， XS-H， and R groups via irregular diet combined with exhaustive swimming for 15 days. The CON group received a standard diet， while other groups were fed a high-fat diet for 10 weeks to establish the hyperlipidemia model. After successful modeling， rats were treated for 8 weeks： M-XSLJZ was administered at 3.51， 7.02， 14.04 g·kg^-1 in XS-L， XS-M， and XS-H groups， respectively. The R group received 9×10^-4 g·kg^-1 of a reference drug. D-xylose excretion rate was measured by the phloroglucinol method. Blood lipids were assessed using an automated biochemical analyzer. Hematoxylin-eosin （HE） staining was used to evaluate the pathological conditions of the liver， and oil red O staining was used to observe the lipid deposition in the liver. The levels of LPS， portal vein serum LPS， LPS-binding protein （LBP）， serum interleukin-6 （IL-6）， IL-1β， and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence was used to evaluate CD86 expression and CD68/TLR4 co-localization in the liver. Protein levels of TLR4， MyD88， NF-κB p65， and p-NF-κB p65 in Kupffer cells were analyzed via Western blot automated protein analysis. Hepatic IL-6， TNF-α， and IL-1β mRNA and protein levels were measured using Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultsCompared with the CON group， the SD-HFD group showed a decrease in D-xylose excretion （P<0.01）. TC， TG， HDL-C， and LDL-C increased （P<0.05， P<0.01）. A large number of hepatic lipid vacuoles and orange-red lipid droplet deposition appeared in the liver. Ileal LPS， portal LPS， and LBP increased （P<0.05， P<0.01）. The levels of serum IL-6， TNF-α， and IL-1β increased （P<0.01）. The expression of CD86 was upregulated （P<0.01）， and the co-expression of CD68 and TLR4 was enhanced. The protein levels of TLR4， MyD88， and p-p65 in Kupffer cells increased （P<0.01）. The mRNA and protein levels of IL-6， TNF-α， and IL-1β increased （P<0.05， P<0.01）. Compared with the HFD group， the SD-HFD group exhibited decreased D-xylose excretion （P<0.01）， higher HDL-C， LDL-C （P<0.05）， increased portal LBP and LPS （P<0.05）， increased serum IL-6 and TNF-α （P<0.01）， upregulated CD86 （P<0.01）， enhanced CD68/TLR4 co-expression， and higher TNF-α mRNA/protein （P<0.05）. Compared with the SD-HFD group， all M-XSLJZ treatment groups showed reduced TC， TG， and LDL-C （P<0.05， P<0.01）. XS-H and R groups displayed improved hepatic lipid deposition. XS-H and R groups had lower ileal LPS， portal LPS， and LBP levels （P<0.05， P<0.01）. All M-XSLJZ treatment groups exhibited reduced serum IL-6， IL-1β， and TNF-α （P<0.01）. The XS-H group showed downregulated CD86 （P<0.01） and weakened CD68/TLR4 co-expression. The XS-H group had reduced TLR4， MyD88， and p-NF-κB p65 in Kupffer cells （P<0.01）. XS-H and R groups showed lower IL-6， TNF-α， and IL-1β mRNA/protein （P<0.05， P<0.01）. ConclusionM-XSLJZ may exert its lipid-lowering effects by inhibiting intestinal-derived LPS and alleviating Kupffer cell inflammation in the liver.

Objective:To observe the clinical progression of low-risk papillary thyroid microcarcinoma（LR-PTMC）, analyze the influencing factors of its oncological outcomes, and explore the feasibility of active surveillance（AS） of LR-PTMC. Methods:This study adopted a prospective observational research design. A total of 85 subjects diagnosed with LR-PTMC during health checkup in Health Management Center of our hospital from March 2021 to October 2022 were enrolled as the research subjects, for at least 2 years of AS follow-up observation. The clinical progress and oncological outcomes were recorded, disease progression was defined as any increase in nodule diameter ≥3 mm or the appearance of new lesions or lymph node metastasis or distant metastasis, and the oncological outcome was use disease progression defining. Cox proportional hazards regression model was used to analyze the influencing factors of oncological outcomes in LR-PTMC patients. Results:A total of 85 LR-PTMC patients who underwent physical examinations were included in this study. The median follow-up time was 2 years, and a total of 23 patients（27.06%） experienced disease progression. Among them, 18 patients（21.18%） had enlarged lesions（any nodule diameter increased by ≥3 mm）, and 5 patients（5.88%） had abnormal or metastatic cervical lymph nodes. The 2-year cumulative disease progression rate was 9.41%. The incidence age of LR-PTMC patients was younger, with a higher proportion of ≤45 years old. The proportion of baseline nodules with a maximum diameter greater than 5 mm is higher, and the proportion of baseline TPO Ab positivity was higher. Ultrasound showed a higher proportion of microcalcifications compared to the non progression group, and the differences were statistically significant（all P<0.05）. Multivariate Cox proportional hazards regression analysis showed that age of onset ≤45 years RR 95% CI 1.052（1.018-1.088） and ultrasound showing microcalcifications RR 95% CI 3.361（1.379-8.194） were independent risk factors affecting disease progression during AS in LR-PTMC patients（P<0.05）. Conclusion:Most LR-PTMC patients maintain stable lesion size and low lymph node metastasis rate during the AS process, with good oncological outcomes in the short term. AS can be considered as a safe and effective alternative to surgical treatment for LR-PTMC patients. But for patients with onset age ≤45 years and microcalcifications, the follow-up interval can be shortened for close observation.
Humans ; Thyroid Neoplasms/pathology* ; Disease Progression ; Prospective Studies ; Carcinoma, Papillary/pathology* ; Female ; Male ; Middle Aged ; Adult ; Watchful Waiting ; Lymphatic Metastasis ; Proportional Hazards Models ; Risk Factors

Cervical chylous leakage is rare after central lymph node dissection for thyroid cancer, and the coexistence of chylothorax is even more uncommon. This article reports a case of a 39-year-old female patient who underwent total thyroidectomy and left central lymph node dissection for papillary thyroid carcinoma complicated by follicular adenoma. On the second postoperative day, the patient developed white, milky drainage from the neck, suggesting chylous leakage, and experienced mild chest tightness, cough, and low-grade fever. Chest CT revealed bilateral pleural effusion, with the left side being more severe. On the third day, ultrasound-guided left-sided pleural puncture and drainage were performed and hydrothorax triglycerides and total cholesterol levels were measured, chylothorax were diagnosed as a result.After conservative treatment including a low-fat diet, fasting, nutritional support, neck pressure dressing and negative pressure suctionand closed chest drainage, the patient's condition improved rapidly. The patient was discharged on the 8th postoperative day with full recovery. No recurrence has been observed during follow-up to date.

Objective To observe the performance and safety of self-developed real-time three-dimensional intracardiac echocardiography(ICE)system(system).Methods The self-developed system was constructed using disposable sterile real-time three-dimensional ICE catheter(catheter)and multifunctional cart-mounted digital ultrasound imaging host(ultrasound host).The diameter of catheter was 10F,with effective length of 90 cm and two-dimensional array transducer array(840 elements)integrated at tip.The ultrasound host was connected to the catheter through matching connector.The performance of this system was evaluated with self-made experimental equipment and standard phantom,and live animal experiment was performed to observe its safety and imaging quality.Results The maximum imaging depth of this system was ≥60 mm.Its axial resolution was ≤1 mm and lateral resolution was ≤1 mm within the depth of 40 mm,the horizontal and vertical geometric position accuracy errors were ≤10％and ≤5％,respectively,while the image geometric distortion was ≤10％,and the measurement volume error was ≤30％.The catheter was successfully inserted into right atrium of pig through femoral vein under ultrasound guidance,smoothly passing through the vascular pathway without any bending or jamming with good controllability.The cardiac images of this system were clear,which completely displayed cardiac chamber structures,and the image resolution met diagnostic requirement.No injury related to interventional procedures was found in laboratory tests nor anatomical results.Conclusion The self-developed ICE system was stable and safe,and initial results showed it could meet clinical application expectations.