The spine is the most common site of skeletal metastasis in lung cancer, which frequently leads to severe complications such as pathological fracture and neurological compromise and is associated with poor prognosis. The development and progression of spinal metastasis from lung cancer are linked to the unique local microenvironment and tumor microenvironment (TME) of the vertebral column. During metastatic evolution, the dense vascular network of the spine and a plethora of signaling molecules, together with the complex cellular constituents and their intricate interactions within the TME, all cooperate to facilitate the tumor invasion and colonization of the vertebral compartment. Mechanistic studies delineating the role of the TME in spinal metastasis from lung cancer have markedly expanded, fostering the emergence of innovative therapeutic strategies—including nanomedicines, sono-photodynamic therapy, gene therapy, and combination regimens. These strategies demonstrate remarkably potential for clinical translation and offer new directions for the precision management of spinal metastasis from lung cancer.

Objective:Hepatocellular carcinoma(HCC)is the most common primary malignant tumor of the liver.The diagnosis rate of early HCC is low,and most patients are diagnosed at the late stage and have a very poor prognosis.Therefore,it is urgent to explore effective early diagnosis markers and intervention targets for HCC.Cancer/testicular antigen 1A(CTAG1A)is abnormally expressed and highly immunogenic in a variety of tumors,but its expression characteristics and immunogenicity in HCC remain unclear.The aim of this study is to identify the expression and immunogenicity of CTAG1A in HCC tissues and cells,providing a new biomarker for the early diagnosis of HCC and a new potential target for clinical immunotherapy.Methods:This study screened the differentially expressed gene profiles between 10 pairs of very early HCC(BCLC stage 0 HCC)tumors and paracancerous tissues using a transcriptome microarray.The expression of CTAG1A was verified by RT-qPCR in an independent large sample(BCLC stage 0,A,B,C HCC tissues and adjacent non-tumor tissues,n=149)and various hepatocellular carcinoma cell lines.Bioinformatics tools(TepiTool of IEDB database and Swiss Model)were used to predict the MHC-Ⅰ and MHC-Ⅱ epitopes of CTAG1A.The candidate peptides were synthesized by solid-phase polypeptide synthesis method.After purification by HPLC and verification by mass spectrometry assay,the specific T cell responses of peripheral blood mononuclear cells(PBMC)of 9 HCC patients to all peptides were detected by IFN-γ enzyme-linked immunospot assay(ELISpot).The clinical samples were collected from HCC patients admitted to the Third Affiliated Hospital of Naval Medical University(Eastern Hepatobiliary Surgery Hospital)from 2015 to 2022.The collection and usage of all samples were carried out with the consent of the patients,and with the approval of the Ethics Committee of Eastern Hepatobiliary Surgery Hospital(EHBHKY2015-01-017)and in strict accordance with relevant requirements and ethical regulations.Statistical analysis was performed using SPSS 30.0 software,and the diagnostic efficiency was evaluated by ROC curve.Results:Transcriptome chip screening results showed that CTAG1A expression was significantly up-regulated in the very early-stage HCC(BCLC stage 0 HCC)(|FC|=99.16,P<0.0001).The verification using the clinical independent samples showed its high expression in all stages of HCC and better diagnostic efficacy in early-stage HCC(BCLC stage 0 HCC AUC=0.6893,sensitivity=85.71%;BCLC stage A HCC AUC=0.8229,sensitivity=83.33%).Furthermore,the expression of CTAG1A was significantly higher in multiple liver cancer cell lines than in relatively normal liver cell lines(P<0.001).Compared with alpha-fetoprotein(AFP),CTAG1A showed better diagnostic efficacy in BCLC stage 0 and stage A HCC(ROC curve analysis of AFP showed no significant difference in early HCC,P>0.05).Bioinformatics tools predicted that CTAG1A contained 8 MHC-type I and 4 MHC-type II epitopes.The IFN-γ ELISpot assay showed that 12 synthetic peptides could induce PBMC specific T cell response in HCC patients to varying degrees.Conclusion:CTAG1A is significantly overexpressed in early-stage HCC and has multi-epitope immunogenicity,which may activate CD8? and CD4? T cells,suggesting its potential as a target for HCC immunotherapy.It may provide a new direction for developing combined immunotherapy strategies based on mRNA vaccines or adoptive cell therapy.Compared with AFP,CTAG1A exhibits better diagnostic efficacy in early-stage HCC,suggesting its potential as a marker for early diagnosis of HCC.

AIM:To investigate the role of specific long noncoding RNA SLC25a6(lncSLC25a6)in homocys-teine(Hcy)-induced cuproptosis in cardiomyocytes.METHODS:Rat cardiomyocytes were cultured in vitro and divided into control group and Hcy group.After 48 h of intervention,the expression levels of cuproptosis-related proteins,ferre-doxin 1(FDX1)and heat shock protein 70(HSP70),were detected by Western blot and immunofluorescence staining.The oxidative stress state of cardiomyocytes was assessed using fluorescence staining,and the intracellular Cu2+levels were measured using a copper ion assay kit.Furthermore,the impact of Hcy on the expression of cuproptosis-related proteins in cardiomyocytes was analyzed following overexpression of lncSLC25a6.RESULTS:Compared with the control group,80 μmol/L Hcy significantly accelerated cardiomyocyte damage,with a notable underexpression of lncSLC25a6(P<0.05).Western blot results indicated that,compared with the control group,the expression level of FDX1 in the Hcy intervention group was significantly reduced(P<0.05),while the expression level of HSP70 was significantly elevated(P<0.05),and the expression level of copper ions in cardiomyocytes of the Hcy group was increased(P<0.05).Immunofluorescence staining showed a significant reduction in FDX1 fluorescence intensity and a significant increase in HSP70 fluorescence in-tensity in the Hcy group.Further overexpression of lncSLC25a6 significantly mitigated Hcy-induced cuproptosis in cardio-myocytes,resulting in elevated expression of FDX1 and reduced expression of HSP70(P<0.05).Pearson correlation analysis demonstrated that the expression level of lncSLC25a6 was negatively correlated with FDX1 protein expression(r=-0.676,P=0.046)and positively correlated with HSP70 expression(r=0.680,P=0.044).CONCLUSION:lnc-SLC25a6 significantly mitigates Hcy-induced cuproptosis in cardiomyocytes,positioning it as a potential therapeutic target for managing Hcy-induced cardiac injury.

The COVID-19 epidemic has spread to the whole world for three years and has had a serious impact on human life, health and economic activities. China's epidemic prevention and control has gone through the following stages: emergency unconventional stage, emergency normalization stage, and the transitional stage from the emergency normalization to the "Category B infectious disease treated as Category B" normalization, and achieved a major and decisive victory. The designated hospitals for prevention and control of COVID-19 epidemic in Tianjin has successfully completed its tasks in all stages of epidemic prevention and control, and has accumulated valuable experience. This article summarizes the experience of constructing a hospital infection prevention and control system during the "Category B infectious disease treated as Category A" period in designated hospital. The experience is summarized as the "Cluster" hospital infection prevention and control system, namely "three rings" outside, middle and inside, "three districts" of green, orange and red, "three things" before, during and after the event, "two-day pre-purification" and "two-director system", and "one zone" management. In emergency situations, we adopt a simplified version of the cluster hospital infection prevention and control system. In emergency situations, a simplified version of the "Cluster" hospital infection prevention and control system can be adopted. This system has the following characteristics: firstly, the system emphasizes the characteristics of "cluster" and the overall management of key measures to avoid any shortcomings. The second, it emphasizes the transformation of infection control concepts to maximize the safety of medical services through infection control. The third, it emphasizes the optimization of the process. The prevention and control measures should be comprehensive and focused, while also preventing excessive use. The measures emphasize the use of the least resources to achieve the best infection control effect. The fourth, it emphasizes the quality control work of infection control, pays attention to the importance of the process, and advocates the concept of "system slimming, process fattening". Fifthly, it emphasizes that the future development depends on artificial intelligence, in order to improve the quality and efficiency of prevention and control to the greatest extent. Sixth, hospitals need to strengthen continuous training and retraining. We utilize diverse training methods, including artificial intelligence, to ensure that infection control policies and procedures are simple. We have established an evaluation and feedback mechanism to ensure that medical personnel are in an emergency state at all times.

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Humans ; Apoptosis Inducing Factor/metabolism* ; NAD/metabolism* ; Dimerization ; Apoptosis

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15％ of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.T260A, p.R422W, and p.R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5％?49.7％ of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3％?17.9％, which was significantly higher than that (6.9％?7.4％) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.

Objective:To investigate the effect of Ziyin Bushen sequential method on the expression of steroids acute regulatory synthesis protein (StAR) and silent information regulator 1 (SIRT1) in ovaries of polycystic ovary syndrome (PCOS) rats.Methods:PCOS rat model was established by dehydroepiandrosterone (DHEA). Eighty 21-day-old clean SD rats were randomly divided into control group (16 rats) and PCOS group (64 rats). PCOS group was further divided into normal saline (NS) group, Ziyin group, Bushen group and Xuguan group with 16 rats in each group. Each group was given intragastric administration for 20 d. Serum estradiol, progesterone and testosterone levels were measured by enzyme linked immunosorbent assay (ELISA) method. The expression levels of StAR, SIRT1 protein and mRNA were detected by Western blotting and RT-PCR.Results:The serum estradiol level in normal saline group was significantly higher than that in control group ( P=0.004). The levels of serum estradiol in Ziyin group, Bushen group and Xuguan group were significantly lower than that in NS group ( P=0.005, P=0.003 and P=0.002, respectively). The serum testosterone level in normal saline group was significantly higher than that in control group ( P=0.005); the level of serum testosterone in Xuguan group was significantly lower than that in NS group, Ziyin group and Bushen group ( P=0.002, P=0.005 and P=0.005, respectively). Compared with control group, the StAR protein content in the normal saline group was higher ( P<0.001), while SIRT1 mRNA was significantly decreased ( P<0.001). Compared with NS group, Ziyin group and Bushen group, the expression of StAR protein and mRNA in Xuguan group was significantly lower ( P<0.001, P=0.008 and P<0.001, respectively); the expression of SIRT1 protein in Xuguan group was significantly higher than that in NS group and Bushen group (all P<0.001); compared with the control group, StAR mRNA in NS group was significantly increased ( P<0.001), while SIRT1 mRNA was significantly decreased ( P<0.001); the expression of SIRT1 gene in Ziyin group and Xuguan group was significantly higher than that in NS group (all P<0.001), while that in Xuguan group was significantly higher than that in Bushen group ( P<0.001). Conclusion:Ziyin Bushen sequential method may decrease the serum androgen level of PCOS rats by upregulating SIRT1 gene expression and StAR in ovary expression.

Objective:To investigate the effect of Ziyin Bushen sequential method on the expression of steroids acute regulatory synthesis protein (StAR) and silent information regulator 1 (SIRT1) in ovaries of polycystic ovary syndrome (PCOS) rats.Methods:PCOS rat model was established by dehydroepiandrosterone (DHEA). Eighty 21-day-old clean SD rats were randomly divided into control group (16 rats) and PCOS group (64 rats). PCOS group was further divided into normal saline (NS) group, Ziyin group, Bushen group and Xuguan group with 16 rats in each group. Each group was given intragastric administration for 20 d. Serum estradiol, progesterone and testosterone levels were measured by enzyme linked immunosorbent assay (ELISA) method. The expression levels of StAR, SIRT1 protein and mRNA were detected by Western blotting and RT-PCR.Results:The serum estradiol level in normal saline group was significantly higher than that in control group ( P=0.004). The levels of serum estradiol in Ziyin group, Bushen group and Xuguan group were significantly lower than that in NS group ( P=0.005, P=0.003 and P=0.002, respectively). The serum testosterone level in normal saline group was significantly higher than that in control group ( P=0.005); the level of serum testosterone in Xuguan group was significantly lower than that in NS group, Ziyin group and Bushen group ( P=0.002, P=0.005 and P=0.005, respectively). Compared with control group, the StAR protein content in the normal saline group was higher ( P<0.001), while SIRT1 mRNA was significantly decreased ( P<0.001). Compared with NS group, Ziyin group and Bushen group, the expression of StAR protein and mRNA in Xuguan group was significantly lower ( P<0.001, P=0.008 and P<0.001, respectively); the expression of SIRT1 protein in Xuguan group was significantly higher than that in NS group and Bushen group (all P<0.001); compared with the control group, StAR mRNA in NS group was significantly increased ( P<0.001), while SIRT1 mRNA was significantly decreased ( P<0.001); the expression of SIRT1 gene in Ziyin group and Xuguan group was significantly higher than that in NS group (all P<0.001), while that in Xuguan group was significantly higher than that in Bushen group ( P<0.001). Conclusion:Ziyin Bushen sequential method may decrease the serum androgen level of PCOS rats by upregulating SIRT1 gene expression and StAR in ovary expression.

Objective To evaluate the postoperative imaging features of lung ultrasound on patients undergoing lung transplantation ,and to provide the evidence for diagnosis and therapy . Methods Between October 2016 and March 2017 ,51 patients undergoing lung transplantation ( unilateral:37 ,bilateral:14 ) admitted to the ICU in Wuxi People′s Hospital were examined by bedside lung ultrasound ,and imaging features were analyzed . Results The main features on ultrasound of 51 patients undergoing lung transplantation were:①Pneumothorax :The A-line arising at the pleural line was shown in all of 51 patients , mainly on anterolateral parts of the chest wall initially ,then fade away towards anterosuperior parts over time . ② Hydrothorax :An anechoic fluid collection was detected ( up to 50 mm in width ) ,and became narrow over time in most patients . A mass of floccules or progressive growth of pleural effusion indicated the need for emergency surgery ,and were confirmed bleeding after surgery . ③Subcutaneous emphysema:The E-line was detected mainly in anterior and lateral parts around the surgical incision of postoperative patients ,and gradually fade away over time . ④Pulmonary edema:On the first postoperative day ,multiple B-lines were shown in 49 cases ,lung consolidation in 36 cases ( mainly in the inferior and inferoposterior parts) ,lung consolidation sonographic air bronchogram in 12 cases . Then the area of consolidation and B-lines reduced ,the air bronchogram sign became more prevalent ,and the shred sign appeared on the border of consolidation over time . Conclusions The imaging features of lung ultrasound provides clinic diagnostic and therapeutic value for postoperative patients undergoing lung transplantation .

Aim To investigate the impact of CYP3 A5 genetic polymorphism on modified releasing tacrolimus pharmacokinetics in Chinese stable renal transplant re-cipients. Methods Pharmacokinetics of once daily-ta-crolimus( tac-q. d. ) and twice daily-tacrolimus( tac-b. i. d. ) were determined by CLIA, CYP3A5 genotype was measured by PCR-RFLP. Each 10 patients receiv-ing tac-q. d. and tac-b. i. d. respectively were en-rolled, and each 5 patients receiving tac-q. d. were matched to poor metabolizer ( PM ) and extensive me-tabolizer ( EM ) group respectively according to CYP3A5 genotypes. Results AUC0~24 h for tac-q. d. was 1. 78 folds higher than AUC0~12 h for tac-b. i. d. , and dose-adjusted C0 was 40% lower for tac-q. d. than for tac-b. i. d. There were no significant differences for other parameters between the two groups; Cmax, AUC0~24 h and C0 were 1. 75, 1. 96 and 2. 49 folds higher for PM than for EM, and dose-adjusted Cmax, AUC0~24 h and C0 were 1. 80, 2. 34 and 2. 64 folds higher for PM than for EM. There were good correla-tions between AUC0~24 h and C0 for tac-q. d. Conclu-sion Conversion from tac-b. i. d. to tac-q. d. results in requirement of increased tacrolimus dose and detec-tion of CYP3A5 genotype, which is necessary for ensu-ring C0 in the range of therapeutic window.