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.

[Objective] To investigate the non-pathological influencing factors of the unqualified alanine aminotransferase (ALT) in the initial screening of blood donors in Changchun and the laboratory re-examination, so as to provide evidence for reducing the deferral of blood donors and the discarding of blood due to ALT disqualification. [Methods] The unqualified results of ALT from the laboratory of our center from September 1, 2023 to October 31, 2024 were collected. The unqualified rates of ALT were statistically analyzed according to the blood collection sites and the initial screening detection equipment. The samples after ALT pre-donation screening were tested in the laborator, and the unqualified rates of ALT in the initial screening and the laboratory, the non-conformity rate of the results and the distribution range of ALT values were statistically analyzed according to the blood collection sites and the initial screening detection equipment. A questionnaire survey was conducted on the blood donors before blood collection to statistically analyze the influence of the blood donors' living habits and diet on ALT test results. [Results] The statistical analysis of the unqualified rate of ALT in the laboratory showed statistically significant differences in the ALT disqualification rates among different blood collection sites and different initial screening detection devices (P<0.05). Comparison of the ALT unqualified rate for the same type of equipment at different sites showed that for Equipment 1, there were differences between the combined blood collection house and the whole blood house, and between the combined blood collection house and the blood donation vehicle (P<0.05); for Equipment 2, there were differences between the combined blood collection house and the blood donation vehicle, and between the whole blood house and the blood donation vehicle (P<0.05); there were no significant differences among other groups with the same equipment. The initial screening and the laboratory test results for the same samples were compared, with unqualified rates of ALT of 16.29% and 13.01%, respectively. There were statistically significant differences in the unqualified rates of ALT among different blood collection sites (P<0.05), but no significant differences in the ALT test results among different detection equipment (P>0.05).. The non-conformity rate between the initial screening and the laboratory results was 5.26%, of which 81.15% (99/122) were unqualified in the initial screening but qualified in the laboratory. There were statistically significant differences in those unqualified in the initial screening but qualified in the laboratory among different blood collection sites and different detection equipment (P<0.05). The median ALT level in the initial screening was 29.0 U/L (with a 5%-95% range of 14-75 U/L), and the median ALT level in the laboratory was 19 U/L (with a 5%-95% range of 8-65 U/L). The results of the questionnaire survey showed that 33.3% (2/6) of those who consumed alcohol within 24 hours before blood donation had unqualified ALT, and 10% (1/10) of those who stayed up late the night before blood donation had unqualified ALT. [Conclusion] The unqualified rates of ALT in the initial screening before blood collection and the laboratory re-examination of blood donors in Changchun are closely related to the blood collection sites, detection equipment, detection environment, detection personnel, samples, ALT thresholds and detection time. Drinking alcohol and staying up late within 24 hours before blood donation increase the risk of unqualified ALT detection.

Objective:To analyze the blood-transition prototype components and metabolites of Fengliaoxing Fengshi Dieda medicinal liquor.Methods:Ultra-high performance liquid chromatographyquadrupole/electrostatic field orbital trap high resolution mass spectrometry (UHPLC-Q Exactive Focus MS/MS) technique was used to compare the chromatogram differences of Fengliaoxing Fengshi Dieda medicinal liquor extract, blank serum and drug-containing serum. According to the retention time, relative molecular weight and the ratio with primary and secondary ion fragments provided by MS, the prototype components and metabolites of Fengliaoxing Fengshi Dieda medicinal liquor extract were analyzed in serum of rats after oral administration. The detection conditions were as follows: the mobile phase of methanol (A)-0.1% formic acid solution (B) for elution gradient (0-5 min, 5%A; 5-60 min, 5%-95%A; 60-65 min, 95%A), the flow rate of 0.3 ml/min, heated electrospray ionization, detection range of m/z 80-1 200, positive and negative ion scanning modes.Results:A total of 31 transitional components were detected in the serum, of which 9 were prototype components and 22 were metabolites. The 9 prototype components were identified as phenylacetaldehyde, baogongteng C/ erycibellin, p-coumaric acid, 5-Hydroxymethylfurfural, quinic acid, paeonol, 3-Hydroxybenzaldehyde, salicylic acid, and isourecumenol. The 22 metabolites mainly consist of 11 organic acid components, 3 indole components, 2 organic phenolic components, 2 alkaloid components, 1 nucleoside component, 1 amino acid component, 1 lactone component, and 1 sulfonic component. The metabolic pathways were mainly glucuronidation, sulfation and others, which by phase Ⅱ metabolism.Conclusion:Organic phenols and organic acids are the main components that enter the body of Fengliaoxing Fengshi Dieda Medicinal Liquor, while alkaloid compounds and organic acid components may be potential active ingredients for its pharmacological effects.

Objective The kinetic parameters of Lin's squeezing-pressing adjustment manipulation were collected for the analysis of its mechanical characteristics,thus to establish a standardized operating procedure to guide clinical teaching of this technique.Methods Ten healthy male trainees(aged 20-30 years)from the Tuina Department of Guangdong Provincial of Chinese Medicine were selected as the subjects.A multi-point thin-film pressure testing system was used to collect mechanical parameters during the operation of Lin's squeezing-pressing adjustment manipulation.The mechanical characteristics were analyzed,and then a mathematical model of time-mechanics curve was constructed.Results(1)The kinetic parameters of Lin's squeezing-pressing adjustment manipulation were as follows:preload force averaged(279.45±19.36)N with a duration of(0.98±0.03)s,the valley value of preload force averaged(137.45±3.59)N,the maximum impact force was(495.56±7.33)N,the impact duration averaged(0.15±0.01)s,the impact velocity averaged(3 183.96±94.76)N/s,and the impulse was(57.16±1.82)N/s.(2)The fitting function of impact force showed large absolute values for both ascending and descending slopes,and the ascending slope was significantly greater than the descending slope,indicating that the Lin's manipulation stressed on rapid outburst of the strength and withdrawal of the strength.(3)One-way ANOVA revealed no statistically significant differences in the preload force and its duration and valley value,the maximum impact force,and impact time among different operators(P＞0.05).Conclusion The analysis of kinetic parameters demonstrates that skilled operators maintain relatively stable mechanical parameters when performing Lin's squeezing-pressing adjustment manipulation.This study provides a preliminary digital analysis of the mechanical characteristics of Lin's bone-setting manipulation in addressing"bone misalignment and tendon displacement",which supplies objective evaluation criteria for the technique.

Objective To investigate the role of high-mobility group AT-hook 2(HMGA2)in osteogenic differentiation of adipose-derived mesenchymal stem cells(ADSCs)and the effect of Hmga2 knockdown for promoting bone defect repair.Methods Bioinformatics studies using the GEO database and Rstudio software identified HMGA2 as a key factor in adipogenic-osteogenic differentiation balance of ADSCs.The protein-protein interaction network of HMGA2 in osteogenic differentiation was mapped using String and visualized with Cytoscape to predict the downstream targets of HMGA2.Primary mouse ADSCs(mADSCs)were transfected with Hmga2 siRNA,and the changes in osteogenic differentiation of the cells were evaluated using alkaline phosphatase staining and Alizarin red S staining.The expressions of osteogenic markers Runt-related transcription factor 2(RUNX2),osteopontin(OPN),and osteocalcein(OCN)in the transfected cells were detected using RT-qPCR and Western blotting.In a mouse model of critical-sized calvarial defects,mADSCs with Hmga2-knockdown were transplanted into the defect,and bone repair was evaluated 6 weeks later using micro-CT scanning and histological staining.Results GEO database analysis showed that HMGA2 expression was upregulated during adipogenic differentiation of ADSCs.Protein-protein interaction network analysis suggested that the potential HMGA2 targets in osteogenic differentiation of ADSCs included SMAD7,CDH1,CDH2,SNAI1,SMAD9,IGF2BP3,and ALDH1A1.In mADSCs,Hmga2 knockdown significantly upregulated the expressions of RUNX2,OPN,and OCN and increased cellular alkaline phosphatase activity and calcium deposition.In a critical-sized calvarial defect model,transplantation of mADSCs with Hmga2 knockdown significantly promoted new bone formation.Conclusion HMGA2 is a crucial regulator of osteogenic differentiation in ADSCs,and Hmga2 knockdown significantly promotes osteogenic differentiation of ADSCs and accelerates ADSCs-mediated bone defect repair in mice.

Objective To investigate the effect of CXXC finger protein 4 (CXXC4) on the proliferation and apoptosis of pancreatic cancer PANC-1 cells.Methods The expression level of CXXC4 in pancreatic canc-er tissues and its relationship with prognosis and clinicopathological stage of the patients were analyzed in on-line databases.The qRT-PCR technique was used to detect the mRNA expression level of CXXC4 in human normal pancreatic ductal epithelial cell line (HPNE) and pancreatic cancer cell PANC-1,AsPC-1 and BxPC-3. si-NC,si-CXXC4,pcDNA3.1 and pCDNA3.1-CXXC4 were respectively transfected into PANC-1 cells.West-ern blot was conducted to detect the effectiveness of CXXC4 knockdown and overexpression.CCK-8,colony formation,EdU and immunofluorescence assays were conducted to analyze the effect of CXXC4 knockdown or overexpression on the proliferation and apoptosis of PANC-1 cells.The bioinformatic websites was used to predict the upstream microRNA (miRNA) of CXXC4.The Starbase database was adopted to analyze the cor-relation between miR-450b-5p and CXXC4 expression in pancreatic cancer tissues.Results The TCGA data-base results showed that the expression of CXXC4 in pancreatic cancer tissues was lowly expressed compared with in paracancerous pancreatic tissues (P<0.001),moreover which was associated with the overall survival and poor prognosis in the patients with pancreatic cancer (P<0.05).The GEPIA database analysis results showed that compared with stage Ⅰ pancreatic cancer,the CXXC4 expression in stage Ⅱ pancreatic cancer was decreased (P<0.05).Compared with HPNE cells,the CXXC4 expression in 3 kinds of pancreatic cancer cells was decreased (P<0.05).Compared with the si-NC group,the proliferation and colony formation ability of PANC-1 cells in the si-CXXC4 group were enhanced,the expressions of proliferation markers Ki67 and PC-NA were increased,and the expressions of apoptosis markers Bax,caspase-3 and caspase-9 were decreased;compared with the pcDNA3.1 group,the PANC-1 cells in the pcDNA3.1-CXXC4 group obtained the opposite results (all P<0.05).The bioinformatic websites predicted that miR-450b-5p was the upstream miRNA of CXXC4,CXXC4 in pancreatic cancer tissues was negatively correlated with the miR-450b-5p expression (r=-0.227) and miR-450b-5p in various mammalian species was highly conserved.Conclusion CXXC4 inhibits the proliferation of PANC-1 cells in pancreatic cancer and promotes theirs apoptosis.

Osteoporotic proximal humeral fracture (OPHF) is one of the common osteoporotic fractures in the aged, with an incidence only lower than vertebral compression fracture, hip fracture, and distal radius fracture. OPHF, secondary to osteoporosis and characterized by poor bone quality, comminuted fracture pattern, slow healing, and severely impaired shoulder joint function, poses a big challenge to the current clinical diagnosis and treatment. In the field of diagnosis, treatment, and rehabilitation of OPHF, traditional Chinese and Western medicine have accumulated rich experience and evidence from evidence-based medicine and achieved favorable outcomes. However, there is still a lack of guidance from a relevant consensus as to how to integrate the advantages of the two medical systems and achieve the integrated diagnosis and treatment. To promote the diagnosis and treatment of OPHF with integrated traditional Chinese and Western medicine, relevant experts from Orthopedic Expert Committee of Geriatric Branch of Chinese Association of Gerontology and Geriatrics, Youth Osteoporosis Group of Orthopedic Branch of Chinese Medical Association, Osteoporosis Group of Orthopedic Surgeon Branch of Chinese Medical Doctor Association, and Osteoporosis Committee of Shanghai Association of Integrated Traditional Chinese and Western Medicine have been organized to formulate Expert consensus on the diagnosis and treatment of osteoporotic proximal humeral fracture with integrated traditional Chinese and Western medicine ( version 2024) by searching related literatures and based on the evidences from evidence-based medicine. This consensus consists of 13 recommendations about the diagnosis, treatment and rehabilitation of OPHF with integrated traditional Chinese medicine and Western medicine, aimed at standardizing, systematizing, and personalizing the diagnosis and treatment of OPHF with integrated traditional Chinse and Western medicine to improve the patients ′ function.

Objective To investigate the role of high-mobility group AT-hook 2(HMGA2)in osteogenic differentiation of adipose-derived mesenchymal stem cells(ADSCs)and the effect of Hmga2 knockdown for promoting bone defect repair.Methods Bioinformatics studies using the GEO database and Rstudio software identified HMGA2 as a key factor in adipogenic-osteogenic differentiation balance of ADSCs.The protein-protein interaction network of HMGA2 in osteogenic differentiation was mapped using String and visualized with Cytoscape to predict the downstream targets of HMGA2.Primary mouse ADSCs(mADSCs)were transfected with Hmga2 siRNA,and the changes in osteogenic differentiation of the cells were evaluated using alkaline phosphatase staining and Alizarin red S staining.The expressions of osteogenic markers Runt-related transcription factor 2(RUNX2),osteopontin(OPN),and osteocalcein(OCN)in the transfected cells were detected using RT-qPCR and Western blotting.In a mouse model of critical-sized calvarial defects,mADSCs with Hmga2-knockdown were transplanted into the defect,and bone repair was evaluated 6 weeks later using micro-CT scanning and histological staining.Results GEO database analysis showed that HMGA2 expression was upregulated during adipogenic differentiation of ADSCs.Protein-protein interaction network analysis suggested that the potential HMGA2 targets in osteogenic differentiation of ADSCs included SMAD7,CDH1,CDH2,SNAI1,SMAD9,IGF2BP3,and ALDH1A1.In mADSCs,Hmga2 knockdown significantly upregulated the expressions of RUNX2,OPN,and OCN and increased cellular alkaline phosphatase activity and calcium deposition.In a critical-sized calvarial defect model,transplantation of mADSCs with Hmga2 knockdown significantly promoted new bone formation.Conclusion HMGA2 is a crucial regulator of osteogenic differentiation in ADSCs,and Hmga2 knockdown significantly promotes osteogenic differentiation of ADSCs and accelerates ADSCs-mediated bone defect repair in mice.

Objective:To explore the clinical value of a trinity strategy for the treatment of multiple orthopedic trauma.Methods:A retrospective case series study was conducted to analyze the clinical data of 1 267 patients with multiple orthopedic trauma admitted to Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine and the First Affiliated Hospital of Navy Medical University from June 2013 to May 2023, including 862 males and 405 females, aged 18-93 years [(55.2±19.8)years]. Associated injuries included hemorrhagic shock in 632 patients, traumatic wet lung in 274, cranial injuries in 135, abdominal and pelvic bleeding in 116, pneumothorax in 89, urinary injury in 13, and vesical rupture in 8. All the patients were treated with the trinity strategy and the treatment process was divided into the phases of first aid, remodeling, and rehabilitation. The first aid phase focused on stabilizing symptoms and saving lives; the remodeling phase centered on restoring the anatomical structure and alignment; the rehabilitation phase aimed for functional recovery through the integration of both Western and traditional Chinese medicine. The all-cause mortality within 30 days after surgery and fracture healing time were calculated; the excellent and good rates of Constant-Murley shoulder score, Mayo elbow score, Gartland-Werley wrist score, Harris hip score, Hospital for Special Surgery (HSS) knee score and the American Orthopedic Foot & Ankle Society (AOFAS) ankle-hindfoot score at the last follow-up and the overall excellent and good rate of all joint function scores were measured. The short form health survey (SF-36) scores were collected preoperatively and at 6 months postoperatively, including 8 aspects such as physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, and mental health. The incidence of postoperative complications was recorded.Results:All the patients were followed up for 6-18 months [(10.2±4.2)months]. The mortality rate during the acute phase (within 30 days after surgery) was 2.37% with 12 deaths due to hemorrhagic shock, 10 due to traumatic brain injury, 6 due to multiple organ dysfunction syndrome (MODS), and 2 due to pulmonary infection. The average fracture healing time averaged 3.8-18 months [(11.5±4.2)months], with 89.49% of the patients having bone union within 12 months after surgery, 8.93% having bone union within 18 months after surgery, and 1.58% undergoing reoperation. For the patients with internal fixation failure and nonunion, the average healing time was extended to (10.2±2.2)months and (13.7±3.3)months respectively. At the last follow-up, the excellent and good rates of Constant-Murley shoulder score, Mayo elbow score, Gartland-Werley wrist score, Harris hip score, HSS knee score, and AOFAS ankle-hindfoot score were 83.93%, 90.24%, 94.12%, 85.57%, 88.46%, and 92.31% respectively, with an overall excellent and good rate of 89.11%. At 6 months after surgery, the SF-36 scores of all the patients in the eight dimensions,including the physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, and mental health were (74.4±8.6)points, (44.7±14.4)points, (77.4±10.9)points, (68.4±18.2)points, (72.5±16.0)points, (76.8±8.7)points, (49.9±17.6)points, and (72.8±17.9)points, significantly improved compared with those before operation [(63.4±12.7)points, (30.9±17.4)points, (56.4±18.0)points, (55.4±24.7)points, (53.5±21.0)points, (55.8±24.3)points, (36.9±24.0)points, (58.8±21.6)points] ( P<0.01). Complications of different degrees occurred in 214 patients (16.89%), including lung infections in 118 patients (9.31%), lower extremity deep vein thrombosis in 50(3.95%), pressure injuries in 26(2.05%), internal fixation failure in 12(0.95%), and nonunion in 8(0.63%). Conclusions:The trinity strategy provides whole-process management, personalized treatment, and overall rehabilitation for multiple orthopedic trauma. It can decrease mortality, shorten fracture healing time, improve joint function and quality of life, and reduce the incidence of complications.