Objective To profile ubiquitination in cysteinyl aspartate-specific proteinase-2(CASP2)deficient cells under heat shock and investigate the role of CASP2 in stress response.Methods Ubiquitination levels in subcellular fractions of control and C ASP2 knockout(KO)cells were detected via Western blotting.After 2 hours of heat shock treatment,Soluble Ⅱ and Pellet fractions were collected from both control and CASP2 KO cells for ubiquitinome analysis.Anti-di-glycine remnant(K-ε-GG)antibody-based proteomic analysis was performed to identify differentially ubiquitinated proteins and associated key signaling pathways.Proteins that displayed significantly upregulated ubiquitination in CASP2 KO cells under heat shock were subjected to His-tag pull-down assays to find out whether CASP2 regulated the ubiquitination of these proteins.Results Under heat shock,CASP2 KO cells displayed significantly higher accumulation of overloaded ubiquitinated conjugates in the Pellet fraction compared to controls.Ubiquitinomics analysis revealed substantial alterations in protein ubiquitination patterns following CASP2 KO.One hundred proteins exhibited significantly elevated ubiquitination levels while 36 proteins had their ubiquitination reduced relative to controls.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis indicated that hyper-ubiquitinated proteins were primarily associated with Huntington disease,Alzheimer disease,bile secretion,carbon metabolism and autophagy.His-tag pull-down assays combined with Western blotting revealed increased ubiquitination of nicotinamide adenine dinucleotide reduced-ubiquinone oxidoreductase 1 beta subcomplex subunit 3(NDUFB3)and autophagy-related protein 9A(ATG9A)in CASP2 KO cells under heat shock.Conclusion Overloaded ubiquitinated conjugates are accumulated due to CASP2 deficiency during heat shock.CASP2 modulates ubiquitination levels through multiple signaling pathways.

Objective To evaluate the efficacy and safety of proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors combined with high-intensity statins in the treatment of hypercholesterolemia. Methods A retrospective study was conducted on patients diagnosed with hypercholesterolemia at the First Affiliated Hospital of Zhengzhou University from June 2023 to December 2024. Patients were divided into an observation group (receiving alirocumab combined with atorvastatin) and a control group (receiving atorvastatin alone) based on their actual treatment regimens. The treatment period was 6 months. Primary outcome measures included lipid parameters such as low-density lipoprotein cholesterol (LDL-C), apolipoprotein B (apoB), and lipoprotein (a), (Lp[a]); carotid ultrasound parameters such as intima-media thickness (IMT), plaque number, plaque area; and inflammatory markers including high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), and tumour necrosis factor-α (TNF-α). Adverse reactions were also recorded. Results A total of 103 patients with hypercholesterolemia were included, with 53 in the observation group and 50 in the control group. After six months of treatment, the observation group demonstrated significantly greater reductions in LDL-C, total cholesterol (TC), triglyceride (TG), and apoB compared to the control group (P ＜ 0.05). The reduction in plaque count and plaque area were also significantly greater in the observation group than in the control group (P＝0.004). The reduction in TNF-α was also greater in the observation group (P＝0.006). There was no statistically significant difference in the incidence of adverse reactions between the two groups. Conclusions PCSK9 inhibitors combined with high-intensity statins can effectively reduce LDL-C levels, improve carotid atherosclerotic plaques and systemic inflammatory status in patients with hypercholesterolemia, and demonstrate good safety in real-world clinical settings.

The monkeypox pandemic has caused two public health emergencies of international concern from 2022 to 2025,causing great impact on the global health system. Early diagnosis and treatment of the monkeypox virus depend on quick, sensitive, and accurate detection methods. The clustered regular interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system, characterized by its unique target gene sequence recognition, and cis-and trans-cleavage activity, has been regarded as the next-generation nucleic acid detection tool in the field of pathogen diagnosis. CRISPR/Cas has been used for detecting monkeypox virus in combination with different amplification and result readout approaches. Although it still faces many challenges, the CRISPR/Cas system is expected to achieve diversified development and provide strong technical support for rapid, accurate, and affordable detection approaches for infectious diseases.

Objective To prepare a variety of quality control(QC)materials for SARS-CoV-2 variants as an addition to the conven-tional SARS-CoV-2 nucleic acid QC products for the laboratory detection of mutant strains by optimizing the preparation and purification process of MS2 phage virus-like particle(VLP)technique,and evaluate their performances.Methods The typical mutation sequence fragments or full length S genes were designed and synthesized according to the genomic information of SARS-CoV-2 variants.Then,they were inserted into the downstream of maturase gene,coat protein and the pac-site of MS2 phage to construct a series of recombi-nant expression vectors.After induced by the prokaryotic expression system,the VLP products were purified through the polyethylenei-mine precipitation,ultrafiltration,nuclease digestion,and gel filtration chromatography.The obtained VLP were validated by the nucle-ic acid electrophoresis,protein electrophoresis,protein concentration determination,and fluorescence PCR,and their performances such as nucleic acid residue and stability were also evaluated.Results A total of 10 kinds of VLP containing the targeted sequences of the gene to be tested were prepared.The length of the foreign sequence wrapped in them ranged from 297 bp to 3 822 bp,which could be combined into a variety of QC materials for the mutation detection of different SARS-CoV-2 variants.The prepared VLP QC materials could not be effectively amplified without nucleic acid extraction or reverse transcription steps during the routine nucleic acid detection.The simulated QC samples remained stable after repeated freeze-thaw cycles.They could be stored stably for 2 months at 25 ℃ and 4 weeks at 37 ℃.Conclusion The established preparation and combined purification process of VLP QC materials can encapsulate vari-ous exogenous nucleic acid sequences with different lengths into the viral coat protein to form VLP,with high production efficiency.The VLP QC products prepared by the above process have stable performance and almost no residual exogenous nucleic acid,which can ef-fectively meet clinical requirements and ensure the quality of laboratory testing.

Objective·To evaluate the feasibility and diagnostic performance of diffusion-relaxation correlation spectroscopic imaging(DR-CSI)in assessing the heterogeneity of benign and malignant head and neck tumors,as well as in identifying occult lymph node metastasis(OLNM).Methods·A prospective study was conducted from January to December 2024 at Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,enrolling patients with suspected head and neck tumors who were scheduled for surgery and had a confirmed pathological diagnosis.All patients underwent preoperative routine head and neck plain and contrast-enhanced magnetic resonance imaging(MRI)examinations,including DR-CSI sequence.Conventional imaging parameters,including maximal diameter(MD),depth of invasion(DOI)for tumors,and MD and short diameter(SD)for lymph nodes,were obtained.Post-processing was performed to obtain apparent diffusion coefficient(ADC),T2 value,and D-T2 spectra for all lesions.The compartment segmentation strategy was optimized based on the spectral peak distribution characteristics of different diseases,and the volume fraction(Vi)of each compartment was obtained.Independent sample t-tests,Mann-Whitney U tests,chi-square tests,or Fisher's exact tests were used to compare intergroup differences in clinical data and imaging metrics.Principal component analysis(PCA)and Adonis analysis were employed to evaluate the discriminative ability of imaging metrics among different subtypes of benign tumors.Receiver operating characteristic(ROC)analysis was used to evaluate the ability of univariate and multivariable models to characterize the malignancy of head and neck squamous cell carcinoma(HNSCC)and identify OLNM.Results·A total of 97 cases were collected,including 28 benign tumors and 69 HNSCCs.Fifteen pathologically confirmed OLNMs and 20 benign lymph nodes(BLNs)were also enrolled.Among the 28 benign tumors,there were 6 cases of pleomorphic adenoma stroma-rich(PA stroma-rich),9 cases of pleomorphic adenoma stroma-poor(PA stroma-poor),9 cases of Warthin's tumor(WT),and 4 cases of basal cell adenoma(BCA).Statistically significant differences were observed in certain imaging parameters(ADC,T2,and DR-CSI Vi)among benign tumor subtypes.PCA analysis demonstrated a strong discriminative ability of imaging parameters in distinguishing pathological subtypes of benign tumors(R2=0.64,P<0.001).Among the 69 HNSCCs,47 were classified as Grade 1(well/moderately well-differentiated)and 22 as Grade 2(moderately/poorly differentiated).Compared to Grade 1,Grade 2 showed lower ADC and higher T2 values,though differences were not statistically significant.As HNSCC malignancy increased,VA4 decreased and VB4 increased significantly.OLNM showed a significant increase in SD and VA4 compared to BLNs.The combination of SD and VA4 for preoperative OLNM identification achieved a diagnostic efficiency of 0.843.Conclusion·DR-CSI can analyze diffusion and relaxation characteristics at the sub-voxel level,offering valuable insights for characterizing benign head and neck tumor subtypes,assessing HNSCC malignancy,and identifying OLNMs.Compared to traditional parameters like ADC or T2,DR-CSI provides enhanced tissue microstructure analysis.

Objective To prepare a variety of quality control(QC)materials for SARS-CoV-2 variants as an addition to the conven-tional SARS-CoV-2 nucleic acid QC products for the laboratory detection of mutant strains by optimizing the preparation and purification process of MS2 phage virus-like particle(VLP)technique,and evaluate their performances.Methods The typical mutation sequence fragments or full length S genes were designed and synthesized according to the genomic information of SARS-CoV-2 variants.Then,they were inserted into the downstream of maturase gene,coat protein and the pac-site of MS2 phage to construct a series of recombi-nant expression vectors.After induced by the prokaryotic expression system,the VLP products were purified through the polyethylenei-mine precipitation,ultrafiltration,nuclease digestion,and gel filtration chromatography.The obtained VLP were validated by the nucle-ic acid electrophoresis,protein electrophoresis,protein concentration determination,and fluorescence PCR,and their performances such as nucleic acid residue and stability were also evaluated.Results A total of 10 kinds of VLP containing the targeted sequences of the gene to be tested were prepared.The length of the foreign sequence wrapped in them ranged from 297 bp to 3 822 bp,which could be combined into a variety of QC materials for the mutation detection of different SARS-CoV-2 variants.The prepared VLP QC materials could not be effectively amplified without nucleic acid extraction or reverse transcription steps during the routine nucleic acid detection.The simulated QC samples remained stable after repeated freeze-thaw cycles.They could be stored stably for 2 months at 25 ℃ and 4 weeks at 37 ℃.Conclusion The established preparation and combined purification process of VLP QC materials can encapsulate vari-ous exogenous nucleic acid sequences with different lengths into the viral coat protein to form VLP,with high production efficiency.The VLP QC products prepared by the above process have stable performance and almost no residual exogenous nucleic acid,which can ef-fectively meet clinical requirements and ensure the quality of laboratory testing.

Objective·To evaluate the feasibility and diagnostic performance of diffusion-relaxation correlation spectroscopic imaging(DR-CSI)in assessing the heterogeneity of benign and malignant head and neck tumors,as well as in identifying occult lymph node metastasis(OLNM).Methods·A prospective study was conducted from January to December 2024 at Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,enrolling patients with suspected head and neck tumors who were scheduled for surgery and had a confirmed pathological diagnosis.All patients underwent preoperative routine head and neck plain and contrast-enhanced magnetic resonance imaging(MRI)examinations,including DR-CSI sequence.Conventional imaging parameters,including maximal diameter(MD),depth of invasion(DOI)for tumors,and MD and short diameter(SD)for lymph nodes,were obtained.Post-processing was performed to obtain apparent diffusion coefficient(ADC),T2 value,and D-T2 spectra for all lesions.The compartment segmentation strategy was optimized based on the spectral peak distribution characteristics of different diseases,and the volume fraction(Vi)of each compartment was obtained.Independent sample t-tests,Mann-Whitney U tests,chi-square tests,or Fisher's exact tests were used to compare intergroup differences in clinical data and imaging metrics.Principal component analysis(PCA)and Adonis analysis were employed to evaluate the discriminative ability of imaging metrics among different subtypes of benign tumors.Receiver operating characteristic(ROC)analysis was used to evaluate the ability of univariate and multivariable models to characterize the malignancy of head and neck squamous cell carcinoma(HNSCC)and identify OLNM.Results·A total of 97 cases were collected,including 28 benign tumors and 69 HNSCCs.Fifteen pathologically confirmed OLNMs and 20 benign lymph nodes(BLNs)were also enrolled.Among the 28 benign tumors,there were 6 cases of pleomorphic adenoma stroma-rich(PA stroma-rich),9 cases of pleomorphic adenoma stroma-poor(PA stroma-poor),9 cases of Warthin's tumor(WT),and 4 cases of basal cell adenoma(BCA).Statistically significant differences were observed in certain imaging parameters(ADC,T2,and DR-CSI Vi)among benign tumor subtypes.PCA analysis demonstrated a strong discriminative ability of imaging parameters in distinguishing pathological subtypes of benign tumors(R2=0.64,P<0.001).Among the 69 HNSCCs,47 were classified as Grade 1(well/moderately well-differentiated)and 22 as Grade 2(moderately/poorly differentiated).Compared to Grade 1,Grade 2 showed lower ADC and higher T2 values,though differences were not statistically significant.As HNSCC malignancy increased,VA4 decreased and VB4 increased significantly.OLNM showed a significant increase in SD and VA4 compared to BLNs.The combination of SD and VA4 for preoperative OLNM identification achieved a diagnostic efficiency of 0.843.Conclusion·DR-CSI can analyze diffusion and relaxation characteristics at the sub-voxel level,offering valuable insights for characterizing benign head and neck tumor subtypes,assessing HNSCC malignancy,and identifying OLNMs.Compared to traditional parameters like ADC or T2,DR-CSI provides enhanced tissue microstructure analysis.

The monkeypox pandemic has caused two public health emergencies of international concern from 2022 to 2025,causing great impact on the global health system. Early diagnosis and treatment of the monkeypox virus depend on quick, sensitive, and accurate detection methods. The clustered regular interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system, characterized by its unique target gene sequence recognition, and cis-and trans-cleavage activity, has been regarded as the next-generation nucleic acid detection tool in the field of pathogen diagnosis. CRISPR/Cas has been used for detecting monkeypox virus in combination with different amplification and result readout approaches. Although it still faces many challenges, the CRISPR/Cas system is expected to achieve diversified development and provide strong technical support for rapid, accurate, and affordable detection approaches for infectious diseases.

Inflammatory bowel disease(IBD)is a chronic and recurrent intestinal inflammatory disease.Interleukin(IL)-22,a member of the IL-10 family,protects the intestinal epithelial barrier and maintains intestinal homeostasis.Multiple studies have demonstrated that IL-22 induces inflammation and promotes inflammatory-cancerous transformation of IBD in different settings.IL-22 is involved in colorectal cancer(CRC)development,and it is a potential anti-cancer therapeutic target.This article reviewed the research progress of IL-22 in IBD.

Inflammatory bowel disease(IBD)is a chronic and recurrent intestinal inflammatory disease.Interleukin(IL)-22,a member of the IL-10 family,protects the intestinal epithelial barrier and maintains intestinal homeostasis.Multiple studies have demonstrated that IL-22 induces inflammation and promotes inflammatory-cancerous transformation of IBD in different settings.IL-22 is involved in colorectal cancer(CRC)development,and it is a potential anti-cancer therapeutic target.This article reviewed the research progress of IL-22 in IBD.