Objective To explore the relationship between circadian rhythm genes and the occurrence, development, prognosis, and tumor microenvironment (TME) of lung adenocarcinoma (LUAD). Methods The Cancer Genome Atlas data were used to evaluate the expression, copy number variation, and somatic mutation frequency of circadian gene sets in LUAD. Gene ontology, Kyoto encyclopedia of genes and genomes, and gene set enrichment analysis were used to explore the potential mechanisms by which circadian rhythm genes affected LUAD progression. Cox regression, least absolute shrinkage and selection operator regression, support vector machine recursive feature elimination, and random forest screened circadian genes and established prognostic models, and on this basis constructed nomogram to predict patients’ 1-, 3-, and 5-year survival rates. Kaplan-Meier survival curves, receiver operating characteristic (ROC) curves, and time-dependent ROC curves were drawn to evaluate the predictive ability of the model, and the external dataset of GEO further verified the prognostic value of the prediction model. In addition, we evaluated the association of the prognostic model with immune cells and immune checkpoint genes. Single cell RNA sequencing (scRNA-seq) analysis was used to explore the molecular characteristics between prognostically relevant circadian genes and different immune cell populations in TME. Results Differentially expressed circadian rhythm genes were mainly enriched in biological processes related to cGMP-PKG signaling pathway, lipid and atherosclerosis, and JAK-STAT signaling pathway. Seven circadian rhythm genes: LGR4, CDK1, KLF10, ARNTL2, RORA, NPAS2, PTGDS were screened out, and a RiskScore model was established. According to the median RiskScore, samples were divided into a high-risk group and a low-risk group. Compared with patients in the low-risk group, patients in the high-risk group showed a poorer prognosis (P<0.001). Immunological characterization analysis showed that there were differences in the infiltration of multiple immune cells between the low-risk group and high-risk group. Most immune checkpoint genes had higher expression levels in the high-risk group than those in the low-risk group, and RiskScore was positively correlated with the expression of CD276, TNFSF4, PDCD1LG2, CD274, and TNFRSF9, and negatively correlated with the expression of CD40LG and TNFSF15. The scRNA-seq analysis showed that RORA and KLF10 were mainly expressed in natural killer cells. Conclusion The prognostic model based on seven feature circadian rhythm genes has certain predictive value for predicting survival of LUAD patients. Dysregulated expression of circadian genes may regulate the occurrence, progression as well as prognosis of LUAD through affecting TME, which provides a possible direction for finding potential strategies for treating LUAD from the perspective of mechanism by which circadian disorder affects immune cells.

Objective To determine the prognostic biomarkers and new therapeutic targets of the lung adenocarcinoma (LUAD), based on which to establish a prediction model for the survival of LUAD patients. Methods An integrative analysis was conducted on gene expression and clinicopathologic data of LUAD, which were obtained from the UCSC database. Subsequently, various methods, including screening of differentially expressed genes (DEGs), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and Gene Set Enrichment Analysis (GSEA), were employed to analyze the data. Cox regression and least absolute shrinkage and selection operator (LASSO) regression were used to establish an assessment model. Based on this model, we constructed a nomogram to predict the probable survival of LUAD patients at different time points (1-year, 2-year, 3-year, 5-year, and 10-year). Finally, we evaluated the predictive ability of our model using Kaplan-Meier survival curves, receiver operating characteristic (ROC) curves, and time-dependent ROC curves. The validation group further verified the prognostic value of the model. Results The different-grade pathological subtypes' DEGs were mainly enriched in biological processes such as metabolism of xenobiotics by cytochrome P450, natural killer cell-mediated cytotoxicity, antigen processing and presentation, and regulation of enzyme activity, which were closely related to tumor development. Through Cox regression and LASSO regression, we constructed a reliable prediction model consisting of a five-gene panel (MELTF, MAGEA1, FGF19, DKK4, C14ORF105). The model demonstrated excellent specificity and sensitivity in ROC curves, with an area under the curve (AUC) of 0.675. The time-dependent ROC analysis revealed AUC values of 0.893, 0.713, and 0.632 for 1-year, 3-year, and 5-year survival, respectively. The advantage of the model was also verified in the validation group. Additionally, we developed a nomogram that accurately predicted survival, as demonstrated by calibration curves and C-index. Conclusion We have developed a prognostic prediction model for LUAD consisting of five genes. This novel approach offers clinical practitioners a personalized tool for making informed decisions regarding the prognosis of their patients.

Lung adenocarcinoma (LUAD) is a global malignancy with significant chemoresistance impacting patient prognosis. The pro-tumorigenic role of hyaluronan-mediated motility receptor (HMMR) in LUAD is recognized. This study was designed to investigate the underlying mechanisms by which HMMR affects chemoresistance in LUAD. Bioinformatics presented the expression patterns of HMMR in LUAD patients and the association between HMMR levels and patient survival, followed by qRT-PCR to verify HMMR expression in LUAD tissues and cells. Further, bioinformatics was leveraged to identify the signaling pathways enriched by HMMR and its relevance to glycolytic genes, we also analyzed changes in the glycolytic activity of LUAD cells by manipulating HMMR expression. Stemness was evaluated through cell aggregation assays and Western blot, and drug responsiveness was gauged using CCK-8 assays, alongside flow cytometry for apoptosis analysis. HMMR was highly expressed in LUAD tissues and cells, and this overexpression correlated with poorer prognoses in patients. GSEA showed that HMMR was notably enriched in the glycolysis and gluconeogenesis pathways, correlating positively with the expression of key glycolytic genes. Cellular experiments confirmed that HMMR knockdown notably suppressed aerobic glycolysis in LUAD cells. Moreover, overexpression of HMMR could further enhance the stemness and cisplatin resistance of LUAD cells by stimulating glycolysis. In brief, this study has validated that high levels of HMMR in LUAD are predictive of poor patient prognosis, and that overexpression of HMMR can catalyze aerobic glycolysis, thus promoting stemness and chemoresistance in LUAD cells. Thus, HMMR could be a target for improving chemosensitivity in LUAD.

Aim To predict and verify the potential mechanism of the compatibility of"ginseng-astragalus-pueraria"in protecting islet morphology and improving insulin resistance by using network pharmacology.Methods The active ingredients and targets of the horn medicine were obtained from three platforms:TC-MSP,TCMIP,and BATMAN.The targets of type 2 dia-betes mellitus(T2DM)were obtained from three plat-forms:TTD,OMIM,and disgeNET.The PPI network was constructed by using the STRING database and Cy-toscape 3.9.1;GO and KEGG analysis were per-formed;POCASA 1.1 was used to predict protein binding sites,and AutoDock Vina1.1.2 was used for docking and experimental verification.Results"Gin-seng-astragalus-pueraria"screened out 2 021 targets,of which 152 were closely related to T2DM,and 10 key genes and the AGE-RAGE signaling pathway were i-dentified.Molecular docking showed that quercetin had good binding to RAGE,INS,and PI3K.Experi-ments showed that the horn drug increased insulin binding rate and secretion index and reduced serum in-sulin level and insulin resistance index.These data benefited from"ginseng-astragalus-pueraria"reducing the expression of AGE-RAGE,activating PI3K-Akt,in-hibiting NF-κB,and reducing the expression of IL-6,IL-1β and TNF-α.Conclusion The study suggests that"ginseng-astragalus-pueraria"regulates the AGE-RAGE/PI3K-Akt/NF-κB signaling pathway,repairs damaged islet morphology,and improves insulin resist-ance.

Objective To explore limb coordination patterns and energy flow strategies during the sit-to-stand(STS)transition in individuals with patellofemoral pain(PFP),so as to provide a theoretical evidence for the pathogenesis of PFP and subsequent formulation of treatment and rehabilitation strategies for PFP patients.Methods A totoal of 36 participants was recruited for the STS test.They were divided into the unilateral PFP group(unilateral group),bilateral PFP group(bilateral group),and control group,based on the number of limbs affected by PFP.An infrared motion capture system and a three-dimensioanl force plate were used for motion capture.Visual 3D and Matlab software were used to calculate the trunk and pelvis angles,angular velocities,linear velocities,and proximal and distal joint forces.Additionally,the angles,torques,and joint forces of the hip,knee,and ankle joints,along with the angular and linear velocities of the thigh and shank,were computed.Coupling angles was used to represent coordination patterns via vector coding;the segmental net energy integration method was used to calculate energy flow within segments at each stage.Results For the coordination pattern at frontal plane,the proximal coordination mode frequency of the pelvis-hip coordination in the flexion momentum phase(FMP)was higher in unilateral group than that in bilateral group(P=0.024).In the momentum transfer phase(MTP),the frequency of in-phase coordination in the trunk-pelvis coordination was higher in unilateral group than that in bilateral group(P=0.023),while the frequency of distal coordination was higher in control group than in that in unilateral group(P=0.032).For the knee-ankle coordination pattern,the frequency of distal coordination in control group was lower than that in unilateral and bilateral groups(P=0.025,P=0.005).In segmental energy flow,during the FMP,the energy output from the pelvis during extension phase(MP)was higher in bilateral group than that in control group(P=0.021).Conclusions PFP affects energy flow patterns and coordination patterns at frontal plane during the STS transition.Individuals in unilateral group may engage in lateral pelvic and ankle movements as a dynamic compensation for patellofemoral joint pressure,whereas individuals in bilateral group appear to increase pelvic region energy output and employ a more complex whole-body coordination pattern to compensate for functional deficits in the knee caused by PFP.

Guided by the cancer toxin theory of TCM master Professor Zhou Zhongying,and absorbing the thoughts of academician Tong Xiaolin's theory of state-target,the pathogenesis of multiple myeloma is summarized as deficiency of healthy qi,bone erosion and marrow damage,cancer toxin accumulation,phlegm and stasis mingling,the combat between healthy qi and evil qi,and dynamic evo-lution;a full-cycle anti-cancer and detoxification prevention and treatment strategy for multiple myeloma is proposed that incorporates the ideas of nourishing deficiency and strengthening healthy qi,preventing cancer and detoxification,resolving phlegm and removing stasis throughout the entire treatment process.The scientific connotation of the pathogenesis theory of"healthy qi deficiency and cancer toxin"in multiple myeloma is explained from multiple aspects such as protein overload and tumor microenvironment,and a prescrip-tion-drug medicine system with Xuanbi Xiaoliu Formula as the core is constructed,which provides new ideas and scientific basis for the clinical diagnosis and treatment plan and full-cycle prevention and control model of multiple myeloma combining Chinese and West-ern medicine.

BACKGROUND:The stiffness of muscle fibers and tendons within skeletal muscles is regulated by the neuromuscular system and remains variable.However,observing the mechanical properties of muscle fibers and tendons during complex multi-joint movements is challenging,and the real-time variation patterns of their stiffness are not yet clear.OBJECTIVE:Taking the open-access simulation data of triceps surae at different running speeds and gait phases as an example,to explore the real-time stiffness change rules of muscle fiber stiffness and tendon stiffness.METHODS:OpenSim simulation results of muscle fiber activation,length,velocity parameters,and tendon length parameters of the triceps surae in five long-distance runners at different running speeds were collected from the Website of Stanford University.The instantaneous slope of the force-length relationship curve of muscle fibers and tendons in the Hill-Zajac muscle model used in the simulation was extracted as the real-time stiffness of the triceps surae muscle fibers and tendons.The temporal changes of stiffness indicators of muscle fibers and tendons during gait were analyzed.RESULTS AND CONCLUSION:The regulation of muscle fiber activation-length-velocity status and tendon strain resulted in the stiffness of muscle fibers and tendons changing in the same trend as the applied force.Compared with lower running speeds,the stiffness of the gastrocnemius muscle fibers was higher in the early support phase at higher running speeds(P ≤ 0.01),and the tendon stiffness of the medial head of the gastrocnemius was higher in the early support phase(P≤ 0.02).The stiffness of the gastrocnemius muscle fibers and tendons was lower from the mid-support to the mid-swing phase(P≤ 0.03),and the stiffness of the soleus muscle fibers was higher during the support phase(P ≤ 0.02).Under all running speeds,the stiffness of the triceps surae muscle fibers and tendons showed a trend of being higher during the support phase than during the pre-swing phase(P ≤ 0.03),and the stiffness of the gastrocnemius muscle fibers and tendons increased again in the late swing phase(P ≤ 0.05).These findings indicate that increasing running speed can increase the stiffness of triceps surae muscle fibers and tendons during the stance phase;when running speed and gait phase change,gastrocnemius and soleus muscles have different patterns of muscle fiber and tendon stiffness changes,whereas gastrocnemius can increase its muscle fiber stiffness and tendon stiffness in the late swing phase through pre-activation phenomenon.

Aim To predict and verify the potential mechanism of the compatibility of"ginseng-astragalus-pueraria"in protecting islet morphology and improving insulin resistance by using network pharmacology.Methods The active ingredients and targets of the horn medicine were obtained from three platforms:TC-MSP,TCMIP,and BATMAN.The targets of type 2 dia-betes mellitus(T2DM)were obtained from three plat-forms:TTD,OMIM,and disgeNET.The PPI network was constructed by using the STRING database and Cy-toscape 3.9.1;GO and KEGG analysis were per-formed;POCASA 1.1 was used to predict protein binding sites,and AutoDock Vina1.1.2 was used for docking and experimental verification.Results"Gin-seng-astragalus-pueraria"screened out 2 021 targets,of which 152 were closely related to T2DM,and 10 key genes and the AGE-RAGE signaling pathway were i-dentified.Molecular docking showed that quercetin had good binding to RAGE,INS,and PI3K.Experi-ments showed that the horn drug increased insulin binding rate and secretion index and reduced serum in-sulin level and insulin resistance index.These data benefited from"ginseng-astragalus-pueraria"reducing the expression of AGE-RAGE,activating PI3K-Akt,in-hibiting NF-κB,and reducing the expression of IL-6,IL-1β and TNF-α.Conclusion The study suggests that"ginseng-astragalus-pueraria"regulates the AGE-RAGE/PI3K-Akt/NF-κB signaling pathway,repairs damaged islet morphology,and improves insulin resist-ance.

Objective To explore limb coordination patterns and energy flow strategies during the sit-to-stand(STS)transition in individuals with patellofemoral pain(PFP),so as to provide a theoretical evidence for the pathogenesis of PFP and subsequent formulation of treatment and rehabilitation strategies for PFP patients.Methods A totoal of 36 participants was recruited for the STS test.They were divided into the unilateral PFP group(unilateral group),bilateral PFP group(bilateral group),and control group,based on the number of limbs affected by PFP.An infrared motion capture system and a three-dimensioanl force plate were used for motion capture.Visual 3D and Matlab software were used to calculate the trunk and pelvis angles,angular velocities,linear velocities,and proximal and distal joint forces.Additionally,the angles,torques,and joint forces of the hip,knee,and ankle joints,along with the angular and linear velocities of the thigh and shank,were computed.Coupling angles was used to represent coordination patterns via vector coding;the segmental net energy integration method was used to calculate energy flow within segments at each stage.Results For the coordination pattern at frontal plane,the proximal coordination mode frequency of the pelvis-hip coordination in the flexion momentum phase(FMP)was higher in unilateral group than that in bilateral group(P=0.024).In the momentum transfer phase(MTP),the frequency of in-phase coordination in the trunk-pelvis coordination was higher in unilateral group than that in bilateral group(P=0.023),while the frequency of distal coordination was higher in control group than in that in unilateral group(P=0.032).For the knee-ankle coordination pattern,the frequency of distal coordination in control group was lower than that in unilateral and bilateral groups(P=0.025,P=0.005).In segmental energy flow,during the FMP,the energy output from the pelvis during extension phase(MP)was higher in bilateral group than that in control group(P=0.021).Conclusions PFP affects energy flow patterns and coordination patterns at frontal plane during the STS transition.Individuals in unilateral group may engage in lateral pelvic and ankle movements as a dynamic compensation for patellofemoral joint pressure,whereas individuals in bilateral group appear to increase pelvic region energy output and employ a more complex whole-body coordination pattern to compensate for functional deficits in the knee caused by PFP.

Objective:To explore the safety and efficacy of intraosseous regional administration (IORA) of vancomycin for preventing infection in primary total knee arthroplasty (TKA).Methods:A total of 124 patients with knee osteoarthritis undergoing TKA between February 2024 and May 2024 at nine hospitals were enrolled. Preoperative infection prophylaxis involved either IORA (0.5 g vancomycin administered via intraosseous regional infusion before incision) or intravenous infusion (1 g vancomycin via peripheral vein). The IORA group included 15 males and 47 females with a median age of 66.5 years (range, 60.0-70.0 years), while the intravenous group included 14 males and 48 females with a median age of 66.0 years (range, 61.8-70.3 years) years. Intraoperative samples were collected including fat and synovium tissues after incision, before prosthesis placement, and after tourniquet release; distal femoral cancellous bone during femoral osteotomy; proximal tibial cancellous bone during tibial osteotomy; proximal intercondylar cancellous bone before prosthesis placement; and peripheral blood from non-infused arms at surgery initiation and after tourniquet release. Vancomycin concentrations were measured using liquid chromatography-tandem mass spectrometry. Vital sign changes were recorded from admission to 5~10 minutes post-IORA (IORA group) or post-incision (intravenous group). Follow-ups were conducted on postoperative day 1 and 3, and at 1 and 3 months, to document complications including IORA-related adverse events, periprosthetic joint infections, surgical site infections, red man syndrome, acute kidney injury, deep vein thrombosis and so on.Results:Vancomycin concentrations in bone, fat, and synovial tissue samples were significantly higher in the IORA group than in the intravenous group ( P<0.05), while vancomycin concentrations in blood samples were significantly lower in the IORA group than in the intravenous group ( P<0.05). Only 7.3%(41/558) of tissue samples in the IORA group had vancomycin concentrations below 2.0 μg/g (the minimum inhibitory concentration of vancomycin against coagulase-negative staphylococcus), compared to 59.3%(331/558) in the intravenous group (χ 2=11.285, P<0.001). In the intravenous group, 16.9%(21/124) of blood samples had vancomycin concentrations exceeding 15.0 mg/L (the threshold associated with a significantly increased risk of nephrotoxicity), while all concentrations in the IORA group were below this threshold, the difference was statistically significant (χ 2=22.943, P<0.001). There were no statistically significant difference ( P>0.05) in vital signs changes before and after vancomycin administration between the two groups. Two patients in the intravenous group experienced incision exudate, while no other related complications occurred in either group. Conclusions:Compared to the traditional intravenous infusion of 1 g vancomycin, intraosseous injection of a low dose (0.5 g) of vancomycin achieves higher local tissue concentrations in the knee joint with a lower incidence of adverse reactions and is safe for infection prophylaxis. Despite guidelines not recommending the routine use of vancomycin for preventing infection after primary TKA, intraosseous injection of 0.5 g vancomycin may be considered intraoperatively for primary TKA in the following scenarios: patients in medical institutions with a high prevalence of methicillin-resistant staphylococcus aureus (MRSA) infections, patients with potential preoperative MRSA colonization, or patients with cephalosporin allergy.