OBJECTIVE: To standardize the operative procedure for tissue-engineered cartilage repair, by demonstrating surgical technique of arthroscopic implantation of decalcified cortex-cancellous bone scaffolds, and summarizing the surgical experience of the sports medicine department team at Peking University Third Hospital. METHODS: This article elaborates on surgical techniques and skills, focusing on the unabridged implantation technology and surgical procedure of decalcified cortex-cancellous bone scaffolds under arthroscopy: First, the patient was placed in the supine position. After anesthesia had been established, the surgeon established an arthroscope and explored the damaged area under the scope. After confirming the size and location of the injury site, the surgeon cleaned the damaged cartilage, and also trimmed the edges of the cartilage to ensure that the cut surface was smooth and stable. the surgeon performed the micro-fracture surgery in the area of cartilage injury, and then measured the size of the injured area under the scope. Next, the surgeon manually trimmed the tissue-engineered scaffold based on the measurements taken under the arthroscope, and then directly implanted the scaffold using a sleeve. A honeycomb-shaped fixator was used to implant absorbable nails to fix the scaffold. After the scaffold was installed, the knee was repeatedly flexed and extended for 10-20 times to ensure stability and range of motion. Finally, the arthroscope was withdrawn and the wound was closed. RESULTS: Decalcified cortex-cancellous bone scaffolds possessed unparalleled advantages over synthetic materials in terms of morphology and biomechanics. The cancellous bone part of the scaffold provided a three-dimensional, porous space for cell growth, while the cortical bone part offered the necessary mechanical strength. The surgery was performed entirely under arthroscopy to minimize invasiveness to the patient. Absorbable pins were used for fixation to ensure the stability of the scaffold. This technique could effectively improve the prognosis of the patients with cartilage injuries and standardized the surgical procedures for arthroscopic tissue-engineered scaffold operations in the patients with cartilage damage. CONCLUSION With the standard arthroscopic tissue-engineered scaffold repair technique, it is possible to successfully repair damaged cartilage, alleviate symptoms in the short term, and provide a more ideal long-term prognosis. The author and their team explain the surgical procedures for tissue-engineered scaffolds under arthroscopy, with the aim of guiding future clinical practice.
Tissue Engineering/methods* ; Humans ; Tissue Scaffolds ; Arthroscopy/methods* ; Cartilage, Articular/surgery*

Approximately 30%-40% of epilepsy patients do not respond well to adequate anti-seizure medications (ASMs), a condition known as pharmacoresistant epilepsy. The management of pharmacoresistant epilepsy remains an intractable issue in the clinic. Its early prediction is important for prevention and diagnosis. However, it still lacks effective predictors and approaches. Here, a classical model of pharmacoresistant temporal lobe epilepsy (TLE) was established to screen pharmacoresistant and pharmaco-responsive individuals by applying phenytoin to amygdaloid-kindled rats. Ictal electroencephalograms (EEGs) recorded before phenytoin treatment were analyzed. Based on ictal EEGs from pharmacoresistant and pharmaco-responsive rats, a convolutional neural network predictive model was constructed to predict pharmacoresistance, and achieved 78% prediction accuracy. We further found the ictal EEGs from pharmacoresistant rats have a lower gamma-band power, which was verified in seizure EEGs from pharmacoresistant TLE patients. Prospectively, therapies targeting the subiculum in those predicted as "pharmacoresistant" individual rats significantly reduced the subsequent occurrence of pharmacoresistance. These results demonstrate a new methodology to predict whether TLE individuals become resistant to ASMs in a classic pharmacoresistant TLE model. This may be of translational importance for the precise management of pharmacoresistant TLE.
Epilepsy, Temporal Lobe/diagnosis* ; Animals ; Drug Resistant Epilepsy/drug therapy* ; Electroencephalography/methods* ; Rats ; Anticonvulsants/pharmacology* ; Neural Networks, Computer ; Male ; Humans ; Phenytoin/pharmacology* ; Adult ; Disease Models, Animal ; Female ; Rats, Sprague-Dawley ; Young Adult ; Convolutional Neural Networks

Aim To investigate the pharmacological mechanism of Ebselenin(Ebselen,EbSe)in the treat-ment of Escherichia coli(E.coli)infection,which had no significant inhibitory effect on Gram-negative bacte-ria,based on previous studies.Methods After EbSe intervention in E.coli infected Raw264.7 cells,the via-bility of Raw264.7 cells was determined by CCK-8 method,the morphology and structure of Raw264.7 cells were observed by electron microscope,and the in-tracellular bacterial load of Raw264.7 cells was calcu-lated by coated plate method.Polarization status of peritoneal macrophages,Raw264.7 intracellular NO and ROS content and intracellular HO-1 expression in Raw264.7 and E.coli acutely infected mice after E.co-li infection by flow cytometry.qPCR was used to detect the expression of related mRNAs in Raw264.7 cells.qPCR was used to detect the intracellular GSH content in Raw264.7 cells by spectrophotometric assay,and the state of cytoskeletal proteins was observed by immuno-fluorescence.Western blot assay was performed to de-tect the intracellular Txnrd1 expression level.Results Microtiter method,CCK-8,and electron microscopy observations showed that EbSe had no effect on the growth of E.coli and Raw264.7 cells in vitro.The re-sults of smear plate counting showed that EbSe reduced the intracellular bacterial load of Raw264.7 in the in-fected group.Flow cytometry results showed that EbSe upregulated the number of M2-type macrophages.The EbSe-treated infected group had reduced intracellular NO and ROS levels and increased GSH levels.The qPCR results showed that the expression of IL-6,IL-1β,and iNOS was decreased,and the expression of HO-1,Txnrd1,and Glut1 was increased in DHB4-in-fected Raw264.7 cells after EbSe treatment.Cytoskel-etal staining showed that the morphology of the EbSe-treated infected cells was similar to that of oxPAPC-in-duced cells.Western blot results showed the expres-sion of Txnrd1 protein in EbSe-treated infected cells in-creased.Conclusion EbSe exerts anti-E.coli acute infection effect by regulating macrophage polarization and inhibiting macrophage excessive inflammatory state.

Objective To investigate the mechanism of Kuijie Ankang Decoction in regulating immune,autophagy and intestinal flora in the treatment of ulcerative colitis(UC).Methods UC mouse model was established by free drinking with sodium dextran sulfate.The mice were randomly divided into blank control group,model group,Kuijie Ankang Decoction group,salazine sulfopyridine(SASP)group and 3-methyladenine(3-MA)group,with 12 mice in each group.Each drug group was given corresponding drugs for gavage,the blank control group and model group were given the same volume of distilled water for gavage for 7 days.The general condition of mice was observed and the disease activity index(DAI)was scored,the morphology of colon tissue was observed by HE staining,the contents of TNF-α,IL-1β,IL-6,IL-8 and IL-10 in colon tissue were detected by ELISA,the mRNA and protein expressions of LC3,Beclin-1 and p62 in colon tissue were detected by RT-qPCR and Western blot,respectively.16S rDNA sequencing was used to analyze the structure of intestinal flora.Results Compared with the blank control group,the mice in the model group showed a decrease in body mass,an increase in DAI score,a decrease in colon length,serious mucosal injury and inflammatory cell infiltration,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue significantly increased(P<0.05,P<0.01),the content of IL-10 decreased(P<0.01),the mRNA expressions of LC3 and Beclin-1 in colon tissue decreased(P<0.05,P<0.01),the mRNA and protein expression of p62 increased(P<0.01),while the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins decreased(P<0.05,P<0.01).Compared with the model group,the body mass of mice in Kuijie Ankang Decoction group and SASP group increased(P<0.05),DAI score decreased(P<0.05),the colon length increased(P<0.05),the pathological damage of colon mucosa was alleviated,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue decreased(P<0.05,P<0.01),the content of IL-10 increased(P<0.01),the expressions of LC3 and Beclin-1 mRNA in colon tissue increased(P<0.05,P<0.01),the expression of p62 mRNA and protein decreased(P<0.05,P<0.01),the expressions of LC3Ⅱ/LC3Ⅰand Beclin-1 protein increased(P<0.05,P<0.01).16S rDNA sequencing results showed that the diversity and evenness of the intestinal flora in the model group mice decreased,with a decrease in the relative abundance of Firmicutes,Actinobacteria and Patescibacteria(P<0.05),and an increase in the relative abundance of Bacteroidota,Verrucomicrobiota and Proteobacteria(P<0.05);the relative abundance of Bacilli and Coriobacteriia decreased(P<0.05),the relative abundance of Bacteroidia,Clostridia and Verrucomicrobiae increased(P<0.05);the relative abundance of Ligilactobacillus and Dubosiella decreased(P<0.05),the relative abundance of unclassified Muribaculaceae,Lachnospiraceae NK4A136_group,Akkermansia and unclassified Lachnospiraceae increased(P<0.05).Compared with the model group,the diversity and evenness of intestinal flora increased in Kuijie Ankang Decoction group and SASP group,with an increase in the relative abundance of Firmicutes(P<0.05),a decrease in the relative abundance of Bacteroidota and Verrucomicrobiota(P<0.05),the relative abundance of Bacteroidia and Bacilli increased(P<0.05),the relative abundance of Verrucomicrobiae decreased(P<0.05);the relative abundance of unclassified Muribaculaceae and Ligilactobacillus increased(P<0.05),while the relative abundance of Lachnospiraceae NK4A136_group and Akkermansia decreased(P<0.05).Conclusion Kuijie Ankang Decoction can significantly improve the intestinal mucosal injury of UC mice,and the mechanism may be related to the regulation of colon autophagy level and intestinal flora disorder.

Objective To investigate the neural basis of glucagon-like peptide-1(GLP-1)in regulating glucose homeostasis and elucidate the molecular mechanisms.Methods Male Glp1r-IRES-Cre,Glp1r-KO,and wild-type mice were used in this study.Fiber photometry was employed to record Ca2+signals of neurons in ventromedial hypothalamus(VMH)and patch-clamp was used to analyze electrophysiological properties of GLP-1 receptor-positive(GLP-1RVMH)neurons.Viral stereotaxic injections,chemogenetics,plasma hormone assays,and routine glucose metabolism assessments were combined to determine the regulatory role of GLP-1RVMH neurons in glucose homeostasis.Tissue and cell mitochondrial respiratory function assays,transmission electron microscopy,and conventional molecular biology methods were used to explore the mechanism by which GLP-1R agonists regulate glucose homeostasis.Results When the glucose concentration decreased from 5.0 mmol/L to 0.5 mmol/L,the action potential frequency of GLP-1RVMH neuron decreased significantly[(4.51±0.80)Hz vs.(1.43±0.51)Hz,P<0.01].Activation of GLP-1RVMH neuron significantly enhanced insulin secretion[(7.60±0.56)μU/mL vs.(11.34±0.93)μU/mL,P<0.01],while inhibition of these neuronal activities impaired the hypoglycemic efficiency of GLP-1 agonists[(32.03%±0.91%)vs.(25.77%±1.09%),P<0.001)].Mechanistically,GLP-1 regulated glucose homeostasis through Drp1 phosphorylation-mediated mitochondrial fission and improved mitochondrial energy metabolism.Conclusion GLP-1RVMH neurons are a class of glucose-excited neurons,and which activated directly promotes secretion of insulin.The hypoglycemic effect of GLP-1R agonists depend on the neuronal activity of GLP-1RVMH.

Objective: To analyze the chemical constituents of Shanxiangyuanye (Turpiniae Folium) through liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, and to evaluate their anti-oxidant, hypoglycemic, and anti-glycation activities related to diabetic complications. Methods: The supernatant of Shanxiangyuanye (Turpiniae Folium) (TFS), obtained following water extraction and alcohol precipitation, was analyzed by LC-MS/MS. Antioxidant activity of TFS in vitro was evaluated using three experimental approaches: the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+) radical cation decolorization assay, and the hydroxyl (·OH) radical scavenging assay. To comprehensively evaluate hypoglycemic potential, α-glucosidase inhibition was measured to analyze in vitro hypoglycemic activity. Subsequently, in vitro models were developed to examine anti-glycation activity through the bovine serum albumin (BSA)-fructose (Fru), BSA-methylglyoxal (MGO), BSA-glyoxal (GO), and D-arginine (Arg)-MGO systems, with particular attention to the inhibitory effects of TFS. Furthermore, the concentrations of fructosamine, protein carbonyls, sulfhydryl groups, and β-amyloid in the glycation solution were quantified using the BSA-Fru model following 7-d of incubation at 37 °C. Results: Using LC-MS/MS analysis in both positive and negative ion modes, we identified 750 chemical components in TFS, primarily including organic acids, amino acids, and their derivatives. In vitro activity studies demonstrated that TFS exhibited remarkable free radical scavenging capacity, with half-maximal inhibitory concentrations (IC50) of 0.47, 1.56, and 0.36 mg/mL against DPPH, ABTS+, and ·OH radicals, respectively. Regarding hypoglycemic activity, TFS dose-dependently inhibited α-glucosidase activity (IC50 = 0.21 mg/mL), displaying comparable efficacy to the clinical drug acarbose (IC50 = 0.23 mg/mL). Notably, TFS intervened in the glycation process: IC50 values were 0.22, 1.91 – 4.96, and 4.09 mg/mL in the BSA-Fru, BSA-MGO/GO, and Arg-MGO models, respectively, with the most prominent inhibitory effects observed in the BSA-Fru model. Furthermore, although TFS may not effectively preserve thiol groups in BSA or reduce thiol oxidation during glycation, it significantly reduces fructosamine levels (in a dose-dependent manner), decreases β-amyloid formation, and inhibits protein carbonylation (P < 0.000 1). Conclusion The findings demonstrate that TFS exhibits a complex chemical composition with potent antioxidant, hypoglycemic, and anti-glycation activities. These results provide compelling scientific evidence supporting TFS’s potential as a natural adjuvant for diabetes prevention and complication management, while laying a solid foundation for its applications in functional food development and adjunctive antidiabetic therapeutics.

Oral squamous cell carcinoma(OSCC)is a common malignant tumor.Interleukin-8(IL-8)is an important biomarker of OSCC,and its level can reflect the occurrence and development of OSCC.It is of great significance to detect IL-8 rapidly and sensitively for the purpose of early diagnosis of OSCC.In this study,gold nanoparticles(AuNPs)with uniform particle size were synthesized by reduction of chloroauric acid with trisodium citrate,and the probe(AuNPs@mAb1)was prepared by coupling AuNPs with the murine anti-IL-8 monoclonal antibody mAb1.Cy5-NHS and murine anti-IL-8 monoclonal antibody mAb2 complex(Cy5-mAb2)and sheep anti-mouse IgG antibody were sprayed on nitrocellulose membrane to form test line(T line)and control line(C line)respectively,and reverse fluorescence-enhanced test strips were thus constructed,based on which an immunochromatographic method was established for highly sensitive detection of IL-8 in saliva samples.The experimental results showed that the test strip had good stability,high specificity and high sensitivity.The linear range for fluorescence detection of IL-8 was 0.01-100 ng/mL,and the limit of detection(3σ)was 7.93 pg/mL.The linear range for visualization detection was 6-100 ng/mL,with limit of detection(3σ)of 0.85 ng/mL.The fabricated test trip had good preparation reproducibility,with inter-and intra-batch assay precision of less than 5.5%.The test strip was used for detection of IL-8 in healthy human saliva samples,with spiked recoveries of 93.7% -102.4%,and relative standard deviations of 2.1% -4.3%.The fabricated test strip could be used for early screening of OSCC.

Objective To develop and validate a risk prediction model for non-cuffed catheter(NCC)dysfunction in hemodialysis patients,aiming to provide a reference for early clinical identification and warning.Methods A prospective study design was adopted.A total of 569 patients with indwelling NCC from the hemodialysis center of a tertiary hospital in Nanchang between December 1,2023 to May 20,2024,were included as a modeling cohort.An additional 172 patients from the hemodialysis center of a tertiary hospital in Changsha,enrolled between May 30 to October 20,2024,formed a validation cohort.Data were collected on general patient characteristics,dialysis information,catheterization details,and clinical parameters.The risk prediction model was constructed using a combination of variables identified through univariate analysis,Lasso regression,logistic regression,and the Boruta algorithm.Model performance was evaluated accordingly.Results The incidence of NCC dysfunction in hemodialysis patients was 44.94％.A total of 5 common predictors were identified by both algorithms,including age,ultrafiltration volume,catheter insertion site,catheter indwelling time,and C-reactive protein.The area under the receiver operating characteristic curve(AUC)was 0.720 for internal validation and 0.766 for external validation.The Brier scores for curve calibration were 0.213 and 0.203,respectively.The decision curve analysis showed clinical benefit within risk threshold ranges of 22％～82％and 22％～96％,respectively.Conclusion The risk prediction model developed in this study demonstrates good predictive performance and can serve as a screening and assessment tool for identifying the risk of NCC dysfunction in hemodialysis patients.

Objective Ticks serve as vectors for transmitting Babesia microti.However,the specific mechanism remains unclear.This study aimed to investigate the effect of tick autophagy molecules on the proliferation of Babesia microti.Methods An experimental model of infected and uninfected mice was used to collect tick materials for proteomic analysis to identify differentially expressed autophagy-related molecules in Haemaphysalis longicornis.The cloning of the HlATG8 gene,protein expression,and production of polyclonal antibodies were completed.The HlATG8 gene was then knocked down using RNAi interference technology.Results The tick autophagy molecule,HlATG8,was identified and found to be significantly upregulated in ticks infected with Babesia microti.The load of Babesia microti in ticks increased significantly following the knockdown of the HlATG8 gene.Conclusions The tick autophagy molecule in Hae.longicornis,HlATG8,inhibits the proliferation of Babesia.

Objective:To explore the value of ultrasound features modified version 2022 of the Ovarian-Adnexal Imaging Reporting and Data System（O-RADS）Category 4 in the differential diagnosis of benign and malignant ovarian-adnexal tumors.Methods:Retrospective analysis was conducted in 501 cases with ovarian masses classified into 4 categories according to the 2022 version of O-RADS who were collected from 4 clinical centers［the Second Afliated Hospital of Harbin Medical University（188 cases），Zhejiang Provincial People's Hospital（146 cases），Sichuan Provincial Maternity and Child Health Care Hospital（90 cases），and Fuling Hospital of Chongqing University（77 cases）］from January 2018 to July 2024 with concomitant surgical resection.The 424 cases from 3 of the clinical centers（the Second Hospital of Harbin Medical University，Zhejiang Provincial People's Hospital，and Sichuan Maternal and Child Health Hospital）were randomly divided into a training group（339 cases）and an internal validation group（85 cases）according to an 8∶2 randomization，while the cases from the other clinical center（Fuling Hospital of Chongqing University）were selected as the external validation group（77 cases），and the pathological diagnosis was used as the “gold standard”.Univariate and multifactorial logistic regression analyses were performed on the ultrasound characteristics of the training group to screen the independent predictors associated with ovarian carcinogenesis，and to formulate the stratification rules for the 4 types of masses in O-RADS. The ROC curve of this stratification method was plotted and the area under the curve（AUC）was calculated，and it was validated in the internal validation group and the external validation group；and the diagnostic accuracy was compared with that of the 2022 version of O-RADS.Results:Univariate logistic analysis showed that cysts with solid components，≥ 4 papillary projections，smooth inner wall of the cyst，color flow score ≥ 3 points，and acoustic shadowing were independent predictors of ovarian cancer（all P < 0.05）；while multifactorial logistic analysis showed that cysts with a solid component and a color flow score ≥3 points were independent risk factors of ovarian cancer（all P < 0.05），and smooth cyst walls and acoustic shadows were independent protective factors（all P < 0.05）.The diagnostic accuracies of the modified training group，internal validation group，and external validation group were 73.7%，68.2%，70.1%，respectively，which were significantly higher than the diagnostic accuracies of the 2022 version of the O-RADS（38.9%，37.6%，33.8%）（all P < 0.05）.The diagnostic sensitivity，specificity and AUC of the training group were 0.871，0.652，0.762，respectively，while the internal validation group were 0.844，0.585，0.714，and 0.846，0.627，0.737 in the external validation group. Conclusions:Improvement of the 2022 version of O-RADS category 4 using ultrasound features may improve the identification of benign and malignant ovarian-adnexal tumors.