Objective:To investigate the impact of postoperative complications on adverse outcomes following curative-intent resection for gallbladder cancer (GBC).Methods:The multi-center real-world study was conducted. The clinicopathological data of 629 patients with GBC, who were admitted to 14 medical centers including The First Affiliated Hospital of Army Medical University from the national multicenter database of Biliary Surgery Group of Elite Group of Chinese Journal of Digestive Surgery, from April 2020 to April 2024 were collected. There were 225 males and 404 females, aged (64±10)years. Patients underwent open curative-intent resection for GBC. Observation indicators: (1)surgery, postoperative complica-tions and adverse outcomes; (2) analysis of risk factors affecting postoperative adverse outcomes in patients and population attributable fraction (PAF). Missing data in predictor variables were addressed using multiple imputation with chained equations, while cases with missing outcome variables were addressed using the "multiple imputation then deletion (MID)" strategy. The severity of multicollinearity among independent variables was assessed using the variance inflation factor (VIF) test. Multivariable possion regression models with log link and robust error variance were construc-ted incorporating restricted cubic splines (3 knots) to address nonlinear relationships in continuous variables, calculating adjusted relative risk ( RR) with corresponding 95% confidence interval ( CI). Adjusted PAF was calculated for each imputed dataset using the AF package of R software, with subsequent pooling performed according to Rubin's rules. Results:(1) Surgery, postoperative complications and adverse outcomes. All 629 patients underwent curative-intent resection for GBC, of which 143 cases had postoperative complications, including 68 cases of intra-abdominal ascites, 39 cases of pulmonary infection, 21 cases of bile leakage, 12 cases of intra-abdominal hemorrhage, 11 cases of liver failure, 10 cases of pan-creatic fistula, 10 cases of wound infection, 10 cases of gastroparesis, 7 cases of cholangitis, 7 cases of sepsis. The same patient could have more than one kind of complication. Of 629 patients, there were 19 cases of postoperative 90-day death and 11 cases of missing data, 42 cases with post-operative 90-day reoperation and 7 cases with missing data, 44 cases with postoperative 90-day readmission and 3 cases with missing data, 155 cases with prolonged postoperative hospital stay and 3 cases with missing data. (2) Analysis of risk factors affecting the postoperative adverse outcomes in patients and PAF. Results of multivariate analysis showed that pulmonary infection and liver failure were independent risk factors for postoperative 90-day mortality ( RR=3.74, 12.15, 95% CI as 1.18-11.83, 1.98-74.48, P<0.05). Pulmonary infection demons-trated the highest PAF as 4.61% (95% CI as 3.94%-5.28%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, and intra-abdominal hemorrhage were independent risk factors for post-operative 90-day reoperation ( RR=4.80, 3.62, 3.46, 4.99, 95% CI as 2.49-9.26, 1.42-9.21, 1.34-8.92, 1.55-16.06, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 8.65% (95% CI as 8.22%-9.08%, P<0.05). Intra-abdominal ascites, bile leakage, and liver failure were independent risk factors for postoperative 90-day readmission ( RR=6.20, 3.33, 14.33, 95% CI as 3.21-11.95, 1.33-8.35, 3.72-55.28, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 9.11% (95% CI as 8.85%-9.37%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, liver failure, and wound infection were independent risk factors for prolonged postoperative hospital stay ( RR=2.29, 2.21, 2.26, 2.14, 3.35, 95% CI as 1.63-3.23, 1.41-3.46, 1.32-3.86, 1.11-4.13, 1.70-6.60, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 6.03% (95% CI as 5.71%-6.35%, P<0.05). Conclusion:Pulmonary infection is the most significant risk factor for postoperative 90-day mortality after curative-intent resection for GBC, while intra-abdominal ascites is the most significant risk factor for postoperative 90-day reoperation, postoperative 90-day readmission, and prolonged postoperative hospital stay.

Objective A non-targeted metabolomics analysis of vitreous fluid from patients with proliferative diabetic retinopathy(PDR)is conducted to explore the"metabolic map"of PDR.This approach aims to deepen the understanding of the disease,identify potential biomarkers.Methods From 35 PDR patients and 30 fresh rhegmatogenous retinal de-tachment(RRD)patients,10 PDR patients with vitreoretinal abnormal adhesions were selected as the experimental group(PDR group),and 10 fresh RRD patients were chosen as the control group(RRD group).Using ultra-high-performance liq-uid chromatography-mass spectrometry non-targeted metabolomics technology,the metabolic profiles of vitreous fluid were analyzed to obtain metabolic spectra.One-dimensional and multidimensional statistical methods were used to analyze the differences in metabolites and metabolic pathways between the PDR and RRD groups.Results A total of 165 differential metabolites were identified in the vitreous humor samples of patients in the PDR and RRD groups,these differential metab-olites were significantly enriched in 21 metabolic pathways(P＜0.05),Among these pathways,those with at least 5 differ-ential metabolites include:methionine and cysteine metabolism;glycine,serine,and threonine metabolism;ascorbic acid and aldose metabolism;amino acid biosynthesis;and central carbon metabolism in cancer.Pyruvate,serine,D-2-phospho-glycerate,threonine,phosphoserine,and high serine are present in multiple metabolic pathways,the areas under the curve are 0.96,0.82,0.85,0.78,0.40,and 0.31,respectively.Conclusion There are 21 significantly different metabolic pathways between PDR and RRD patients.Pyruvate stands out in multiple pathways,potentially serving as a biomarker for PDR diagnosis.

Objective It investigates the current status of negative defensive medical behavior tendencies among medical students and explores influencing factors,aiming to provide evidence for improving medical students' propensity for negative defensive medical practices.Methods In February 2023,a cluster sampling approach was employed to recruit 297 medical students from a medical college-affiliated hospital in Province H.Descriptive statistics,chi-square tests,and correlation analyses were conducted to assess negative defensive medical behavior tendencies.Structural equation modeling(SEM)was utilized to explore multidimensional influencing factors.Results 45％of medical students demonstrated high-level negative defensive medical behavior tendencies.Statistically significant differences were observed between genders(x2=5.569,P＜0.05)and only-child/non-only-child groups(x2=6.104,P＜0.05).Positive correlations were identified between negative defensive medical behaviors and role stress(r=0.217,P＜0.01),negative organizational environment evaluation(r=0.395,P＜0.01),and negative social environment evaluation(r=0.405,P＜0.01).Only-child status was confirmed as an influencing factor(β=-0.123,P=0.036).Negative organizational environment evaluation(β=0.504,P＜0.001)and negative social environment evaluation(β=0.193,P=0.039)showed significant positive effects.Conclusion Immediate interventions-including role-specific risk allocation,organizational ecosystem reconstruction,and social environment governance-require urgent implementation to address early-stage behavioral deviance,counteract the solidification of defensive medical thinking patterns,and reduce predispositions toward negative professional behaviors among medical trainees.

Objective To evaluate the efficacy and safety of Qixian Tongluo Formula in the treatment of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome,and to preliminarily explore the molecular mechanism of Qixian Tongluo Formula in improving impaired motor function from the perspective of cross-kingdom regulation of Chinese medicine microRNA(miRNA).Methods A pragmatic randomized controlled trial was conducted with 102 patients in the recovery period of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome in our hospital.Patients were randomly divided into trial group and control group,with 51 cases in each group.The control group received standard Western medicine standard treatment,while the trial group received Qixian Tongluo Formula in addition to the standard treatment,with one dose per day,boiled in water,and taken warm after breakfast and dinner for a course of 2 months.The disability rate was used as the main efficacy indicator,and the incidence of adverse reactions was used as a safety indicator.miRNA from patient serum and Qixian Tongluo decoction were extracted respectively,and high-throughput sequencing was performed.The two sequences were compared to screen out the cross-kingdom gene transfer of Chinese medicine miRNA.Finally,its target genes of miRNA were predicted,and GO function and KEGG pathway enrichment analysis were carried out.Results A total of 67 patients completed the clinical trial,including 36 cases in the trial group and 31 cases in the control group;The disability rate in the trial group(13.9%)was lower than that in the control group(35.5%)(P<0.05);The incidence of adverse reactions was similar between the trial group(7.69%)and the control group(6.06%)(P>0.05);A total of 9530 Qixian Tongluo decoction miRNA sequences were screened,with 150 potentially involved in cross-kingdom gene transfer,including families such as miR-15 and miR-17;According to the target gene prediction of the top 10 miRNAs in cross-kingdom gene transfer of Chinese medicine,345 overlapping target genes were obtained;GO functional enrichment analysis revealed 16 biological processes,7 cellular components,and 2 molecular functions among the top 25 enriched functions,while KEGG pathway analysis mainly focused on the transforming growth factor-βsignaling pathway,neurotrophin signaling pathway,which are closely related to neural repair and functional recovery processes such as glial scar formation and synaptic plasticity after cerebral ischemia.Conclusion Qixian Tongluo Formula can significantly improve the functional independence level of patients with kidney deficiency and blood stasis syndrome in the recovery period of paralysis after cerebral infarction,offering a safe and effective treatment option for these patients;There were a large number of miRNAs in Qixian Tongluo decoction,some of which could cross-kingdom transferred into the human blood circulation,and promote the recovery of motor function in patients with cerebral infarction through multi-target,multi link and multi pathway gene network regulation.This study provides a new idea for subsequent clinical and basic research.

Objective:To investigate the impact of postoperative complications on adverse outcomes following curative-intent resection for gallbladder cancer (GBC).Methods:The multi-center real-world study was conducted. The clinicopathological data of 629 patients with GBC, who were admitted to 14 medical centers including The First Affiliated Hospital of Army Medical University from the national multicenter database of Biliary Surgery Group of Elite Group of Chinese Journal of Digestive Surgery, from April 2020 to April 2024 were collected. There were 225 males and 404 females, aged (64±10)years. Patients underwent open curative-intent resection for GBC. Observation indicators: (1)surgery, postoperative complica-tions and adverse outcomes; (2) analysis of risk factors affecting postoperative adverse outcomes in patients and population attributable fraction (PAF). Missing data in predictor variables were addressed using multiple imputation with chained equations, while cases with missing outcome variables were addressed using the "multiple imputation then deletion (MID)" strategy. The severity of multicollinearity among independent variables was assessed using the variance inflation factor (VIF) test. Multivariable possion regression models with log link and robust error variance were construc-ted incorporating restricted cubic splines (3 knots) to address nonlinear relationships in continuous variables, calculating adjusted relative risk ( RR) with corresponding 95% confidence interval ( CI). Adjusted PAF was calculated for each imputed dataset using the AF package of R software, with subsequent pooling performed according to Rubin's rules. Results:(1) Surgery, postoperative complications and adverse outcomes. All 629 patients underwent curative-intent resection for GBC, of which 143 cases had postoperative complications, including 68 cases of intra-abdominal ascites, 39 cases of pulmonary infection, 21 cases of bile leakage, 12 cases of intra-abdominal hemorrhage, 11 cases of liver failure, 10 cases of pan-creatic fistula, 10 cases of wound infection, 10 cases of gastroparesis, 7 cases of cholangitis, 7 cases of sepsis. The same patient could have more than one kind of complication. Of 629 patients, there were 19 cases of postoperative 90-day death and 11 cases of missing data, 42 cases with post-operative 90-day reoperation and 7 cases with missing data, 44 cases with postoperative 90-day readmission and 3 cases with missing data, 155 cases with prolonged postoperative hospital stay and 3 cases with missing data. (2) Analysis of risk factors affecting the postoperative adverse outcomes in patients and PAF. Results of multivariate analysis showed that pulmonary infection and liver failure were independent risk factors for postoperative 90-day mortality ( RR=3.74, 12.15, 95% CI as 1.18-11.83, 1.98-74.48, P<0.05). Pulmonary infection demons-trated the highest PAF as 4.61% (95% CI as 3.94%-5.28%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, and intra-abdominal hemorrhage were independent risk factors for post-operative 90-day reoperation ( RR=4.80, 3.62, 3.46, 4.99, 95% CI as 2.49-9.26, 1.42-9.21, 1.34-8.92, 1.55-16.06, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 8.65% (95% CI as 8.22%-9.08%, P<0.05). Intra-abdominal ascites, bile leakage, and liver failure were independent risk factors for postoperative 90-day readmission ( RR=6.20, 3.33, 14.33, 95% CI as 3.21-11.95, 1.33-8.35, 3.72-55.28, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 9.11% (95% CI as 8.85%-9.37%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, liver failure, and wound infection were independent risk factors for prolonged postoperative hospital stay ( RR=2.29, 2.21, 2.26, 2.14, 3.35, 95% CI as 1.63-3.23, 1.41-3.46, 1.32-3.86, 1.11-4.13, 1.70-6.60, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 6.03% (95% CI as 5.71%-6.35%, P<0.05). Conclusion:Pulmonary infection is the most significant risk factor for postoperative 90-day mortality after curative-intent resection for GBC, while intra-abdominal ascites is the most significant risk factor for postoperative 90-day reoperation, postoperative 90-day readmission, and prolonged postoperative hospital stay.

Objective A non-targeted metabolomics analysis of vitreous fluid from patients with proliferative diabetic retinopathy(PDR)is conducted to explore the"metabolic map"of PDR.This approach aims to deepen the understanding of the disease,identify potential biomarkers.Methods From 35 PDR patients and 30 fresh rhegmatogenous retinal de-tachment(RRD)patients,10 PDR patients with vitreoretinal abnormal adhesions were selected as the experimental group(PDR group),and 10 fresh RRD patients were chosen as the control group(RRD group).Using ultra-high-performance liq-uid chromatography-mass spectrometry non-targeted metabolomics technology,the metabolic profiles of vitreous fluid were analyzed to obtain metabolic spectra.One-dimensional and multidimensional statistical methods were used to analyze the differences in metabolites and metabolic pathways between the PDR and RRD groups.Results A total of 165 differential metabolites were identified in the vitreous humor samples of patients in the PDR and RRD groups,these differential metab-olites were significantly enriched in 21 metabolic pathways(P＜0.05),Among these pathways,those with at least 5 differ-ential metabolites include:methionine and cysteine metabolism;glycine,serine,and threonine metabolism;ascorbic acid and aldose metabolism;amino acid biosynthesis;and central carbon metabolism in cancer.Pyruvate,serine,D-2-phospho-glycerate,threonine,phosphoserine,and high serine are present in multiple metabolic pathways,the areas under the curve are 0.96,0.82,0.85,0.78,0.40,and 0.31,respectively.Conclusion There are 21 significantly different metabolic pathways between PDR and RRD patients.Pyruvate stands out in multiple pathways,potentially serving as a biomarker for PDR diagnosis.

Objective To explore the early diagnostic value of serum heat shock protein 70(HSP70),periosten combined with low-dose spiral CT(LDCT)for lung cancer.Methods From July 2022 to June 2024,103 lung cancer patients admitted to our hospital were regarded as the lung cancer group,and 87 patients with benign pulmonary nodules were selected as the benign group.ELISA kit was used to measure serum HSP70 and Periostin.Multivariate logistic regression was applied to analyze the influencing factors of lung cancer.Four grid table method was applied to calculate the sensitivity,specificity,and accuracy of LDCT combined with serum HSP70 and Periostin in the diagnosis of lung cancer.Kappa test was applied to evaluate the consistency between LDCT,serum HSP70,Periostin combined with LDCT and pathological diagnosis of lung cancer.Results Compared with the benign group,the lung cancer group had greatly higher levels of serum HSP70 and Periostin(P＜0.05).Compared to the benign group,the lung cancer group had higher proportions of lobulation and spiculation signs(P＜0.05).Multivariate logistic regression showed that HSP70(OR=1.569),Periostin(OR=1.427),lobulation sign(OR=2.015),and spiculation sign(OR=1.946)were all independent risk factors for lung cancer(P＜0.05).The sensitivity,specificity,and accuracy of LDCT in diagnosing lung cancer were 85.44％,88.51％,and 86.84％,respectively;the sensitivity,specificity,and accuracy of serum HSP70 and Periostin combined with LDCT in diagnosing lung cancer were 93.20％,80.46％,and 87.37％,respectively.Kappa test showed that the consistency between serum HSP70,Periostin combined with LDCT had high consistency with pathology in the diagnosis of lung cancer(Kappa value=0.743).Conclusion The serum levels of HSP70 and Periostin are higher in lung cancer patients.The combination of serum HSP70 and Periostin with LDCT has better diagnostic efficacy for lung cancer.

Objective To explore the characteristics of coagulation and immune function and their clinical significance in children with severe adenovirus pneumonia(SAP).Methods A total of 122 children with SAP and 120 children with non-severe adenovirus pneumonia(NSAP)were enrolled and assigned to severe group and non-severe group.The severe group was further subdivided into good prognosis subgroup(n=105)and poor prognosis subgroup(n=17)according to prognosis.Addition-ally,120 healthy children undergoing physical examination during the same period were included as control group.Coagulation function indicators,including prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(FIB),and thrombin time(TT)were measured.Levels of immunoglobulin A(IgA),immunoglobulin G(IgG),and immunoglobulin M(IgM)were determined using the nephelometric immunoassay.Multivariate Cox regression analysis was conducted to explore the influencing factors of poor prognosis.Results Compared with the control group,children in both the non-severe and severe groups exhibited increased levels of white blood cells(WBC),neutrophils(N),C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),interleukin-6(IL-6),procalci-tonin(PCT),FIB,and IgG,as well as decreased levels of B lymphocytes,natural killer(NK)cells,PT,APTT,IgA,and IgM(P＜0.05).Compared with the non-severe group,children in the severe group showed increased levels of WBC,N,CRP,ESR,IL-6,PCT,FIB,and IgG,as well as decreased levels of B lymphocytes,NK cells,PT,APTT,IgA,and IgM(P＜0.05).Compared with the good prognosis subgroup,the poor prognosis subgroup exhibited increased levels of CRP,IL-6,PCT,and IgG,as well as decreased levels of APTT,IgM,and IgA(P＜0.05).The results of multivariate Cox regression analysis showed that PCT,APTT,and IgM were influencing factors of poor prognosis in children with SAP(P＜0.05).Conclusion Children with SAP have decreased levels of PT,APTT,IgA,and IgM,as well as increased levels of FIB and IgG.Coagulation and im-mune function indicators are correlated with the prognosis of SAP.

Objective To explore the early diagnostic value of serum heat shock protein 70(HSP70),periosten combined with low-dose spiral CT(LDCT)for lung cancer.Methods From July 2022 to June 2024,103 lung cancer patients admitted to our hospital were regarded as the lung cancer group,and 87 patients with benign pulmonary nodules were selected as the benign group.ELISA kit was used to measure serum HSP70 and Periostin.Multivariate logistic regression was applied to analyze the influencing factors of lung cancer.Four grid table method was applied to calculate the sensitivity,specificity,and accuracy of LDCT combined with serum HSP70 and Periostin in the diagnosis of lung cancer.Kappa test was applied to evaluate the consistency between LDCT,serum HSP70,Periostin combined with LDCT and pathological diagnosis of lung cancer.Results Compared with the benign group,the lung cancer group had greatly higher levels of serum HSP70 and Periostin(P＜0.05).Compared to the benign group,the lung cancer group had higher proportions of lobulation and spiculation signs(P＜0.05).Multivariate logistic regression showed that HSP70(OR=1.569),Periostin(OR=1.427),lobulation sign(OR=2.015),and spiculation sign(OR=1.946)were all independent risk factors for lung cancer(P＜0.05).The sensitivity,specificity,and accuracy of LDCT in diagnosing lung cancer were 85.44％,88.51％,and 86.84％,respectively;the sensitivity,specificity,and accuracy of serum HSP70 and Periostin combined with LDCT in diagnosing lung cancer were 93.20％,80.46％,and 87.37％,respectively.Kappa test showed that the consistency between serum HSP70,Periostin combined with LDCT had high consistency with pathology in the diagnosis of lung cancer(Kappa value=0.743).Conclusion The serum levels of HSP70 and Periostin are higher in lung cancer patients.The combination of serum HSP70 and Periostin with LDCT has better diagnostic efficacy for lung cancer.

Objective:To explore the effect of hairy and enhancer of split related protein 2 (Hey2) on osteoclast differentiation through the activation of nuclear factor of activated T cells cytoplasmic 1 (NFATc1).Methods:RAW264.7 cells were induced with receptor activator of NF-κB ligand (RANKL) to differentiate into osteoclasts. Experimental groups were divided by different concentrations of RANKL (0, 10, 20, 50 μg/L) and different processing time (0, 3, 5, 7 days). Hey2 overexpression experiment was grouped as follows: blank control group, RANKL group, empty plasmid vector control group (Hey2-NC+RANKL), Hey2 overexpression group (Hey2-OE+RANKL); similarly, groups in Hey2 knockdown experiment were as follows: blank control group, RANKL group, negative control group (si-NC+RANKL), Hey2 knockdown group (si-Hey2+RANKL). Chromatin immunoprecipitation experiment groups were divided as non-specific IgG control group (IgG control group), non-specific IgG group (IgG RANKL group), Hey2-specific antibody control group (anti-Hey2 control group), Hey2-specific antibody group (anti-Hey2-RANKL group). For the different RANKL concentration groups and different induction time groups, real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the mRNA expressions of nuclear factor of NFATc1, cathepsin K (CTSK), and cellular feline osteosarcoma oncogene (c-Fos) and tartrate-resistant acid phosphatase (TRAP) staining was used to assess the formation of multinucleated osteoclasts. After Hey2 overexpression or knockdown, RT-qPCR and Western blotting were used to detect the gene and protein expressions of NFATc1, c-Fos, and CTSK. TRAP staining was used to evaluate the formation of multinucleated osteoclasts. Bioinformatics prediction (NCBI, JASPAR) and chromatin immunoprecipitation (ChIP) assay were used to validate the binding of Hey2 to the NFATc1 promoter region.Results:During the osteoclastic differentiation of RAW 264.7 cells induced by RANKL, the expression of Hey2 could be detected, and the expression level of Hey2 decreased with the increase of RANKL concentration and induction time. In the 50 μg/L RANKL group, the expression levels of Hey2 gene (0.18±0.00) and protein (0.22±0.02) were significantly lower than those in the control group (1.00±0.00, 0.52±0.01) ( t=41.67, 12.88; both P<0.001). In the 50 μg/L RANKL group inducted for 5 days, the expression levels of Hey2 gene (0.27±0.02) and protein (0.79±0.01) were significantly lower than those in the control group (1.00±0.00, 1.15±0.02) ( t=11.47, 108.60; both P<0.001). Hey2 overexpression significantly reduced the gene and protein expressions of NFATc1, c-Fos, and CTSK, as well as the production of TRAP-positive cells (all P<0.05). Hey2 knockdown significantly increased the gene and protein expressions of NFATc1, c-Fos, and CTSK, as well as the production of TRAP-positive cells (all P<0.05). After inducing RAW264.7 cells with 50 μg/L RANKL for 1 day, ChIP results showed that among the two sample groups treated with Hey2 antibody, the detection level of the NFATc1 promoter region (-400 to -200 bp) in the anti-Hey2-RANKL group (18.06±0.06) was significantly higher than that in the anti-Hey2 control group (13.37±0.36) ( t=12.56, P<0.001). Conclusions:Hey2 can bind to the downstream target gene NFATc1 at -400 to -200 bp region of the promoter. As a transcriptional repressor, Hey2 inhibits osteoclast differentiation.