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.

ObjectiveTo screen and evaluate the antidepressant compounds of Curcumae Rhizoma， and explore its mechanism of regulating the nuclear factor erythroid 2-related factor 2（Nrf2）/glutathione（GSH） peroxidase 4（GPX4）/GSH pathway from an antioxidant perspective. MethodsThe antioxidant activities in vitro of 11 characteristic components from Curcumae Rhizoma， including curcumol， curgerenone， curdione， curzerene， curcumenol， curcumenone， dehydrocurdione， isocurcumenol， furanodienone， furanodiene and zederone， were detected using 1，1-diphenyl-2-picrylhydrazyl（DPPH） and 2，2'-azinobis-（3-ethylbenzothiazoline-6-sulphonic acid） diammonium salt（ABTS） radical scavenging assays. The depression in Drosophila melanogaster was induced by chronic unpredictable mild stress（CUMS）， and W1118 wild-type male D. melanogaster were randomly divided into blank group， model group， curcumol group， curgerenone group， curdione group， curzerene group， curcumenol group，curcumenone group， dehydrocurdione group， isocurcumenol group， furanodienone group， furanodiene group， zederone group and fluoxetine group（10 μmol·L^-1）. The treatment groups received a dose of 0.1 g·L^-1 of 11 characteristic components from Curcumae Rhizoma， while the blank and model groups were administered equivalent volumes of solvent. The sucrose preference test， climbing test and forced swimming test were used to evaluate the behavioral indicators of depression in D. melanogaster. Liquid chromatography-mass spectrometry（LC-MS） was used to detect the levels of 5-hydroxytryptamine（5-HT） and dopamine（DA） in the brain of D. melanogaster， and the entropy weight method was used to comprehensively evaluate neurobehavioral and neurotransmitter indicators， resulting in the identification of the antidepressant active components of Curcumae Rhizoma. In addition， a mouse depression model was established by CUMS， and C57BL/6J mice were randomly divided into blank group， model group， low and high dose groups of curzerene（0.5， 1 mg·kg^-1）， and fluoxetine group（10 mg·kg^-1） to confirm the antidepressant effect of the optimal active ingredient by behavioral analysis. Flow cytometry was used to detect the content of reactive oxygen species（ROS） in the hippocampus of mice from each group. Enzyme-linked immunosorbent assay was used to detect the contents of adenosine triphosphate（ATP）， superoxide dismutase（SOD）， catalase（CAT） and GSH. Transmission electron microscope（TEM） was used to observe the effect of curzerene on the ultrastructure of mitochondria in hippocampal tissue. Western blot was performed to determine the level of Nrf2 protein， and Nrf2 inhibitor（ML385） was used to verify the relationship between the antidepressant effect of curzerene and regulation of Nrf2. Real time fluorescence quantitative polymerase chain reaction（Real-time PCR） was employed to detect the effect of curzerene on the mRNA expression level of GPX. ResultsIn vitro antioxidant experiments showed that curzerene and curgerenone exhibited the most significant ability to scavenge free radicals， and comprehensive evaluation results of entropy weight method indicated that curzerene stood out as the most promising active component. Compared with the blank group， the model group exhibited a significant decrease in sucrose preference coefficient and the number of times entering the open field center（P<0.01）， as well as a significant increase in immobility time in the forced swimming and tail suspension tests（P<0.01）， and the ROS content in hippocampus significantly elevated（P<0.01）， while the ATP content significantly reduced（P<0.01）. In the hippocampal neurons of the model group， mitochondrial cristae were disordered， with vacuolation of the inner membrane and severe damage. Nrf2 protein expression level in the model group was significantly decreased（P<0.05）， and the antioxidant enzymes SOD， CAT and GSH contents were also significantly reduced（P<0.05， P<0.01）， and the gene expression levels of GPX1， GPX4 and GPX7 were significantly decreased（P<0.01）. Compared with the model group， the high-dose group of curzerene showed a significant increase in the sucrose preference coefficient and the number of times entering the open field center（P<0.05）， as well as a significant decrease in immobility time in the forced swimming and tail suspension tests（P<0.05， P<0.01）. The ROS content in the hippocampus of the high-dose group of curzerene was significantly reduced（P<0.01）， while the ATP content was significantly increased（P<0.05）. The neuronal mitochondrial damage in the hippocampus of the high-dose group of curzerene was alleviated， and the expression level of Nrf2 protein was significantly increased（P<0.05）. The Nrf2 inhibitor ML385 reversed the improvement of curzerene on depressive behaviors in CUMS mice. The GSH content in the hippocampal neurons of the high-dose group of curzerene was significantly increased（P<0.01）， while there were no significant differences in SOD and CAT contents. The expression level of GPX4 gene in the hippocampal neurons of the high-dose group of curzerene was significantly increased（P<0.05）， while there were no significant differences in other GPX genes. ConclusionCurzerene is the best component with antidepressant activity in Curcumae Rhizoma. It may improve mitochondrial dysfunction to exert its antidepressant effect by regulating Nrf2 and its downstream GPX4/GSH pathway rather than CAT or SOD pathways.

OBJECTIVE: To explore the genetic etiology of a Chinese pedigree with recurrent Joubert syndrome with negative results by whole-exome sequencing in the prior proband. METHODS: Chinese pedigree which opted elective abortion at the Women and Children's Hospital Affiliated to Chongqing Medical University in December 2024 was selected as the study subject. Whole-genome sequencing was carried out on fetal tissue after termination of pregnancy. Candidate variants were validated by Sanger sequencing and interpreted, while non-coding variant was analyzed using in silico prediction tools. RNA sequencing and cDNA sequencing were conducted on fetal brain tissue. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.2024YL045-02). RESULTS: Both the fetus and the affected child were found to harbor compound heterozygous variants of the CEP290 gene, namely c.7341dup (p.Leu2448fs*8) (pathogenic, maternally inherited) and c.1523-408G>A (likely pathogenic, paternally inherited). Both in silico analysis and fetal brain RNA sequencing confirmed aberrant RNA splicing caused by the intronic variant. CONCLUSION This case has highlighted the value of combining whole-genome sequencing with RNA functional validation. Above results not only enriched the spectrum of CEP290 gene mutations but also underscored its diagnostic value in resolving complex prenatal cases, providing critical clues for the prenatal diagnosis and recurrence risk assessment in genetic counseling.
Female ; Humans ; Pregnancy ; Abnormalities, Multiple/genetics* ; Antigens, Neoplasm/genetics* ; Cell Cycle Proteins/genetics* ; Cerebellum/abnormalities* ; Cytoskeletal Proteins/genetics* ; Eye Abnormalities/genetics* ; Introns/genetics* ; Kidney Diseases, Cystic/diagnosis* ; Pedigree ; Retina/abnormalities* ; Sequence Analysis, RNA/methods* ; Whole Genome Sequencing/methods* ; Child

Objective:To explore differences in the clinical and pathological features and postoperative survival after radical resection of biliary tract cancer in different locations such as intrahepatic cholangiocarcinoma,perihilar cholangiocarcinoma,distal cholangiocarcinoma,and gallbladder cancer.Methods:This is a retrospective case series study. The clinical and pathological data of 4 852 patients with biliary tract cancer admitted to the Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xi ′an Jiaotong University from January 2013 to December 2022 were retrospectively analyzed. Among them, 2 110(43.49%%) patients were male and 2 742(56.51%) patients were female,aged from 26 to 88 years with age of (61.3±10.8) years. Observation indicators: (1) The distribution,diagnosis and treatment of patients with biliary tract cancer; (2) Comparison of clinical and pathological features of patients with biliary tract cancer after curative-intent resection; (3) Survival analysis of patients with biliary tract cancer after curative-intent resection; (4) Analysis of effect on adjuvant therapy for patients with biliary tract cancer after curative-intent resection. One-way analysis of variance,Kruskal-Wallis H test and χ 2 test were used for among-group comparisons,respectively. Survival univariate analysis was performed using the Kaplan-Meier method and Log-rank test. Results:Among the 4 852 patients with biliary tract cancer,there were 2 303 cases (47.46%) of gallbladder cancer,952 cases (19.62%) of intrahepatic cholangiocarcinoma,892 cases (18.38%) of perihilar cholangiocarcinoma,and 705 cases(14.53%) of distal cholangiocarcinoma. From the perspective of the year of diagnosis and treatment,the overall number of patients diagnosed and treated for biliary tract cancer has shown an upward trend. From the perspective of diagnosis and treatment,the curative-intent resection rate was 33.37%(1 619/4 852),and the curative-intent resection rate of distal cholangiocarcinoma was higher than that of other biliary tract cancer ( χ2=23.897, P<0.01). Univariate analysis showed that there were statistical differences in gender,age,bile duct stones,total bilirubin at admission,carcinoembryonic antigen,CA19-9,CA125,the degree of pathological differentiation,vascular invasion,microvascular invasion,perineural invasion,surgical margins,pT staging,and pN staging among patients for biliary tract cancer in different locations (all P<0.05). Survival comparison analysis showed that recurrence-free survival and overall survival of patients with gallbladder cancer after curative-intent resection were significantly better than those of intrahepatic cholangiocarcinoma,perihilar cholangiocarcinoma,and distal cholangiocarcinoma ( χ 2=87.780,83.717,both P<0.01). Comparing the postoperative prognosis of patients with biliary tract cancer between the two periods of 2013 to 2017 and 2018 to 2022, the results showed that recurrence-free survival and overall survival of patients with biliary tract cancer from 2018 to 2022 were significantly better than those from 2013 to 2017 ( χ 2=31.202,25.615, both P<0.01),and the proportion of early recurrence and short-term death after curative-intent resection was significantly reduced ( χ 2=21.588,9.623, both P<0.01),with gallbladder cancer being the most significant ( P<0.01). Postoperative adjuvant therapy for patients with biliary tract cancer can effectively prolong recurrence-free survival and overall survival ( χ 2=5.033,11.273,both P<0.05). Conclusions:Gallbladder cancer remains the most common biliary tract cancer with a relatively favorable prognosis after radical resection. There are significant differences in the clinical and pathological features of biliary tract cancer in different locations,and patients with adjuvant therapy effectively improving prognosis.

Objective To investigate the inhibitory effect of Scutellaria baicalensis combined with 5-Fu on Lewis tumor bearing mouse lung cancer based on the JAK2/STAT3 signaling pathway.Methods To detect the changes in body mass,food intake and tumor volume of Lewis tumor-bearing mice after intervention of Scutellaria baicalensis(20 mg/kg)combined with 5-Fu.Pathological changes in tumor tissue were observed by HE staining,expression levels of proliferation related protein Ki67 were observed by immunohistochemical staining,changes in cell apoptosis levels in tumor tissue were observed by TUNEL staining,and changes in expression levels of JAK2,p-JAK2,STAT3,and p-STAT3 proteins in tumor tissue were observed by immunohistochemical staining.Results Compared with the model group,the tumor volume of mice after combined intervention significantly decreased(P＜0.01)and body mass increased(P＜0.05),but there was no significant change food intake.The expression of proliferation related protein Ki67 in tumor tissue was significantly reduced,and the number of apoptotic cells labeled with TUNEL was significantly increased;The expression of p-JAK2 and p-STAT3 proteins is elevated.Conclusions Scutellaria baicalensis decoction can inhibit the JAK2/STAT3 signaling pathway and increase the inhibitory effect of 5-Fu on mouse lung cancer.

Objective: To investigate whether fibroblast growth factor 18(FGF18) can induce human gingival fibroblasts(HGFs) isolatedin vitroto differentiate into osteoblast-like cells, and to explore the mechanism of osteogenesis. Methods : HGFs were isolated, cultured and identified by tissue block method. The third generation of HGFs were divided into experimental group and control group. FGF18 and L-DMEM was added to the experimental group while L-DMEM was added to the control group.The effects of different concentrations of FGF18(0, 0.01, 0.02, 0.04, 0.06 mg/L) on proliferation of HGFs were detected by Methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay. Alkaline phosphatase(ALP) and alizarin red staining were used to detect the osteogenesis and mineralization ability of the cells after induction. RT-PCR, immunocytochemistry staining, and Western blot were used to detect the expression of genes and proteins related to osteogenesis and BMP2 in the BMP signaling pathway. Results: Compared with the control group, the experimental group could promote the proliferation of HGFs at 3, 5, 7, 9, and 11days(P<0.05),ALP activity and mineral salt deposition increased after induction at 14 and 21 days(P<0.05), and the expressions of ALP, OPN, OCN mRNA and BMP2 mRNA in BMP signaling pathway significantly increased(P<0.01). The expressions of OPN, OCN and BMP2 protein at 21 days were significantly higher than those at 14 days(P<0.01). Conclusion FGF18 can promote the proliferation of HGFs, and induce the differentiation of HGFs into functional osteoblasts. The osteogenic mechanism is related to the upregulation of BMP2.

Objective To explore the clinical efficacy of dual plasma molecular adsorption(DPMAS)sequential plasma exchange(PE)artificial liver mode in the treatment of liver failure(LF).Methods Eighty-five patients with LF receiving the artificial liver treatment in the Affiliated Hospital of Zunyi Medical Univer-sity from January 2020 to December 2023 were selected as the study subjects and divided into the study group(n=52)and the control group(n=33)according to the different treatment modes.The study group conduc-ted DPMAS sequential PE treatment and the control group underwent the PE treatment.The liver function[total bilirubin(TBIL),alanine aminotransferase(ALT),aspartate aminotransferase(AST),serum albumin(ALB),globulin(GLO),prealbumin(PAB)],Hb,coagulation function[platelet(PLT),plasminogen activity(PTA),international normalized ratio(INR),fibrinogen(FIB)]before treatment and at 24 h after treatment were compared between the two groups.Results Compared with before treatment,the levels of TBIL,ALT,AST,GLO and Hb after the first and second treatment in the two groups were decreased,ALB level in the control group and PAB level after the second time treatment was increased(P＜0.05).Compared with after the first treatment,the levels of TBIL,ALT and GLO after the second treatment in the two groups and the levels of AST and Hb in the study group were decreased,ALB level in the study group and PAB level in the two groups were increased(P＜0.05).Compared with before treatment,the levels of PLT and FIB after the first treatment in the two groups and INR level in the control group were decreased,PTA level in the control group was increased(P＜0.05).Compared with before treatment,the levels of PLT,INR and FIB after the second treatment in the two groups were decreased,PTA level was increased(P＜0.05).Compared with be-fore treatment,the levels of PLT,INR and FIB after the second treatment in the two groups were decreased,and PTA level was increased(P＜0.05).Compared with after the first treatment,PTA level after the second treatment in the study group was increased and INR level was decreased.Conclusion PE and DPMAS sequen-tial PE all could improve the liver function in the patients with LF,moreover the two times treatment has more significant effect.

Objective To evaluate the effect of five-element music combined with sandplay therapy in improving negative emotions and enhancing quality of life in patients with malignant tumors.Methods A to-tal of 112 malignant tumor patients meeting inclusion and exclusion criterias were enrolled and randomly di-vided into four groups by using random number table method:control group,five-element music group,sand-play therapy group,and combined therapy group,with 28 patients in every group.The control group received routine nursing and psychological care;the five-element music group received routine nursing,psychological care and five-element music therapy;the sandplay therapy group received routine nursing,psychological care,and sandplay therapy;the combined therapy group received routine nursing,psychological care,five-element music,and sandplay therapy.Interventions were conducted once a week for 6 weeks.Changes in self-rating anxiety scale(SAS),self-rating depression scale(SDS),and quality of life core scale(QLQ-C30)scores were compared among the four groups.Results Before treatment,there were no significant differences in SAS,SDS,or QLQ-C30 subscale scores among the four groups(P>0.05).After treatment,SAS and SDS scores decreased,while QLQ-C30 subscale scores increased in all four groups(P<0.05).After treatment,inter-group comparisons showed significant differences in SAS,SDS,and QLQ-C30 subscale scores(P<0.05),with the combined therapy group showing the best efficacy.The SAS and SDS scores in the five-element music group,sandplay therapy group,and combined therapy group were lower than those in the control group,while QLQ-C30 subscale scores were higher(P<0.05).No significant differences were observed between the five-element music group and sandplay therapy group in SAS,SDS,or QLQ-C30 subscale scores(P>0.05).The combined therapy group had significantly lower SAS and SDS scores and higher QLQ-C30 subscale scores than the five-element music group and sandplay therapy group(P<0.05).Conclusion Five-element music com-bined with sandplay therapy can significantly improve negative emotions and quality of life in patients with malignant tumors.

Objective To analyze the main epidemiological characteristics of fungal infection in this hospital in the past 7 years,and to provide reference for clinical treatment and prevention and control strategies of fun-gal infection.Methods The fungal data and clinical data of related patients isolated from clinical samples in Peking Union Medical College Hospital from early January 2018 to the end of May 2024 were selected,and the main epidemiological characteristics of fungal infection in this hospital were identified and described through multi-angle statistical analysis.Results A total of 4 479 patients with filamentous fungal infection were en-rolled.The proportion of male patients[57.5％(2 576/4 479)]was higher than that of female patients[42.5％(1 903/4 143)],mainly distributed in internal medicine,Intensive Care Unit(ICU)and emergency de-partment,among which internal medicine accounted for the highest proportion[50.0％(2 241/4 479)].About 90.0％of the specimens were from the lower respiratory tract,in addition to specimens from skin and soft tis-sue,tissue,ear and blood culture.In terms of seasonal distribution,there are more patients in winter.The fun-gi were mainly composed of Aspergillus,Mucor,Cerdosporium,Fusarium and Penicillium,among which As-pergillus was the most abundant,accounting for 74.6％of the total.Aspergillus fumigatus was the most a-bundant Aspergillus,accounting for 42.5％of the total Aspergillus(1 418/3 340).Among the related infec-tions caused by mold,Aspergillus was the most common in the lower respiratory tract,accounting for 76.8％.Among them,Aspergillus fumigatus accounted for the highest proportion(33.6％).98.6％of the molds infected the ear were Aspergillus,of which Aspergillus niger and Aspergillus terreus were the most common.Skin infections are mainly caused by Sporothrix schenckii,Trichophyton rubrum,Microsporum ca-nis.The results of in vitro drug sensitivity test showed that the four common Aspergillus isolated in this hos-pital were sensitive to voriconazole,and amphotericin B had better antifungal activity against Mucorales in vitro.Conclusion Based on the main epidemiological characteristics of fungal infections in this hospital,it is recommended that special attention be paid to the admission of patients in the respiratory department during the peak infection period in autumn and winter.In the treatment of fungal infections in different regions and on different body parts,attention should be paid to the differences in the distribution of bacterial species.

Objective:To design protein binders of human epidermal growth factor receptor-2(HER2,also known as ErbB2)using de novo protein design and to preliminarily evaluate their biological effects on tumor cells.Methods:Based on de novo protein design strategy,RosettaDesign and ProteinMPNN algorithms were used to design the binding proteins targeting ErbB2.Concurrently,AlphaFold was applied to assess structural accuracy of the protein binders and their interactions with ErbB2.Subsequently,yeast surface display technology combined with dual-fluorescence high-throughput flow cytometry were performed to screen for protein binders which were capable of specifically binding to ErbB2 specifically.Selected binding proteins were expressed and purified in Eschrichia coli system.Bio-layer interferometry(BLI)was used to validate the binding specificity of the purified protein binders to ErbB2.Finally,Western blotting and CCK-8 assay were used to evaluate the effects of the candidate binding protein on the downstream signaling of ErbB2 and cell proliferation capacity in ovarian cancer(SK-OV-3)and pancreatic cancer(BxPC-3)cell lines,respectively.Results:In this study,the protein binders targeting ErbB2 were successfully designed,a corresponding binding protein library was established,and a protein binder that specifically binds to ErbB2 was screened out from the library.After BLI verification,this protein binder could effectively bind to ErbB2.Further investigation revealed that in ErbB2-high-expressing SK-OV-3 and BxPC-3 cells,this protein binder could effectively inhibit cell proliferation and reduce the phosphorylation level of AKT,a signaling molecule downstream of ErbB2.Conclusion:Based on the strategy of de novo protein design,this study successfully constructed a protein binder that can effectively bind to ErbB2.The binder can effectively inhibit the activation of the downstream signaling pathway mediated by ErbB2 and the proliferation of tumor cells.This indicates the potential application value of de novo designed protein binders targeting ErbB2 in cancer therapy,providing a novel research approach for the field of targeted tumor therapy.