Myocardial fibrosis is a serious cause of heart failure and even sudden cardiac death. However, the mechanisms underlying myocardial ischemia-induced cardiac fibrosis remain unclear. Here, we identified that the expression of sterile alpha and TIR motif containing 1 (SARM1), was increased significantly in the ischemic cardiomyopathy patients, dilated cardiomyopathy patients (GSE116250) and fibrotic heart tissues of mice. Additionally, inhibition or knockdown of SARM1 can improve myocardial fibrosis and cardiac function of myocardial infarction (MI) mice. Moreover, SARM1 fibroblasts-specific knock-in mice had increased deposition of extracellular matrix and impaired cardiac function. Mechanically, elevated expression of SARM1 promotes the deposition of extracellular matrix by directly modulating P4HA1. Notably, by using the Click-iT reaction, we identified that the increased expression of ZDHHC17 promotes the palmitoylation levels of SARM1, thereby accelerating the fibrosis process. Based on the fibrosis-promoting effect of SARM1, we screened several drugs with anti-myocardial fibrosis activity. In conclusion, we have unveiled that palmitoylated SARM1 targeting P4HA1 promotes collagen deposition and myocardial fibrosis. Inhibition of SARM1 is a potential strategy for the treatment of myocardial fibrosis. The sites where SARM1 interacts with P4HA1 and the palmitoylation modification sites of SARM1 may be the active targets for anti-fibrosis drugs.

Deactivation of the mitochondrial pyruvate dehydrogenase complex (PDC) is important for the metabolic switching of cancer cell from oxidative phosphorylation to aerobic glycolysis. Studies examining PDC activity regulation have mainly focused on the phosphorylation of pyruvate dehydrogenase (E1), leaving other post-translational modifications largely unexplored. Here, we demonstrate that the acetylation of Lys 488 of pyruvate dehydrogenase complex component X (PDHX) commonly occurs in hepatocellular carcinoma, disrupting PDC assembly and contributing to lactate-driven epigenetic control of gene expression. PDHX, an E3-binding protein in the PDC, is acetylated by the p300 at Lys 488, impeding the interaction between PDHX and dihydrolipoyl transacetylase (E2), thereby disrupting PDC assembly to inhibit its activation. PDC disruption results in the conversion of most glucose to lactate, contributing to the aerobic glycolysis and H3K56 lactylation-mediated gene expression, facilitating tumor progression. These findings highlight a previously unrecognized role of PDHX acetylation in regulating PDC assembly and activity, linking PDHX Lys 488 acetylation and histone lactylation during hepatocellular carcinoma progression and providing a potential biomarker and therapeutic target for further development.
Humans ; Acetylation ; Carcinoma, Hepatocellular/genetics* ; Liver Neoplasms/genetics* ; Pyruvate Dehydrogenase Complex/genetics* ; Gene Expression Regulation, Neoplastic ; Animals ; Mice ; Cell Line, Tumor ; Protein Processing, Post-Translational ; Histones/metabolism* ; Disease Progression

Gene synthesis is an enabling technology that supports the development of synthetic biology. The existing approaches for de novo gene synthesis generally have tedious operation, low efficiency, high error rates, and limited product lengths, being difficult to support the huge demand of synthetic biology. The assembly and error correction are the keys in gene synthesis. This study first designed the oligonucleotide sequences by reasonably splitting the virus genome of approximately 10 kb by balancing the parameters of sequence design software ability, PCR amplification ability, and assembly enzyme assembly ability. Then, two-step PCR was performed with high-fidelity polymerase to complete the de novo synthesis of 3.0 kb DNA fragments, and error correction reactions were performed with T7 endonuclease Ⅰ for the products from different stages of PCR. Finally, the virus genome was assembled by 3.0 kb DNA fragments from de novo synthesis and error correction and then sequenced. The experimental results showed that the proposed method successfully produced the DNA fragment of about 10 kb and reduced the probability of large fragment mutations during the assembly process, with the lowest error rate reaching 0.36 errors/kb. In summary, this study developed an efficient de novo method for synthesizing a viral genome of about 10 kb with T7 endonuclease Ⅰ-mediated error correction. This method enabled the synthesis of a 10 kb viral genome in one day and the correct plasmid of the viral genome in five days. This study optimized the de novo gene synthesis process, reduced the error rate, simplified the synthesis and assembly steps, and reduced the cost of viral genome assembly.
Genome, Viral/genetics* ; Polymerase Chain Reaction/methods* ; DNA, Viral/genetics* ; Bacteriophage T7/enzymology* ; Synthetic Biology/methods*

Objective:To express,purify,prepare antibodies,and analyze the subcellular localization of Toxoplasma gondii thioredoxin 20(Trx20),and to provide the reference for the development of Toxoplasma gondii vaccine.Methods:Bioinformatics-related websites and software were used to perform bioinformatics analysis of the Trx20 protein;specific primers were designed to amplify the target fragment and construct the prokaryotic expression vector;the protein was expressed in vitro and purified;experimental animals were immunized to prepare antibodies;enzyme-linked immunosorbent assay(ELISA)method was used to detect the titer of the polyclonal antibodies;Western blotting method was used to verify the specificity and sensitivity of the antibodies and to determine the natural expression of the protein;immunofluorescence assay(IFA)was used to analyze the subcellular localization of the protein.Results:The bioinformatics analysis results showed that Trx20 protein was a relatively stable hydrophilic protein with a molecular formula of C2172H3412N548O616S20,containing 424 amino acids,a predicted relative molecular mass of 47 700,and a theoretical isoelectric point of 8.55;it was predicted that the protein had one signal peptide,no transmembrane region,contained one domain named"Thioredoxin like Superfamily",and had 35 phosphorylation sites,one N-glycosylation site,and 17 antigenic determinants;in the secondary structure,alpha-helices accounted for 41.51％of the total amino acids,and random coils accounted for 39.86％;the recombinant plasmids pET-28a-Trx20 and pGEX-4T-1-Trx20 were successfully constructed,and the soluble recombinant protein was expressed and purified;polyclonal antibodies were successfully prepared with a titer as high as 1:64 000,and they specifically recognized the endogenous Trx20 protein in Toxoplasma gondii;the subcellular localization results showed that Trx20 protein was widely distributed in the cytoplasm of the parasite.Conclusion:Toxoplasma gondii Trx20 protein is a secretory protein containing phosphorylation/glycosylation modification sites and a thioredoxin domain,and it is localized in the cytoplasm of the parasite.

Objective To investigate the investigation of co-morbidity etiology and prognosis analysis of chronic diseases in the elderly population. Methods The data of 1 475 elderly patients who were seen and treated in Chengdu Fifth People's Hospital from January 2019 to December 2021 were screened to analyze their disease status, co-morbidity combinations and patterns, co-morbidity influencing factors, and prognosis. Results The top four prevalence rates among 1 475 elderly patients with chronic diseases were hypertension 555 (37.63%), gastric or gastrointestinal diseases 445 (30.17%), arthritis or rheumatism 427 (28.95%), and diabetes 329 (26.58%). 1034 co-morbidities were found in 1475 elderly patients with chronic diseases, with a co-morbidity rate of 70.10%. The binary disease combination accounted for 58.41% and the ternary disease combination accounted for 41.59%. Female, age >70 years, family history of chronic diseases, overweight/obesity, daily physical inactivity, history of alcohol/smoking, poor sleep quality, and poor dietary habits were the independent influencing factors for co-morbidity in elderly patients with chronic diseases (OR_female=2.413, ORage ≥ 70=1.670, OR_{history of alcohol consumption}family history of chronic diseases=2.846, OR_{history of alcohol consumption}overweight/obesity=2.570, ORdaily inactivity=1.802, OR_{history of alcohol consumption}=3.543, ORhistory of smoking=1.784, OR_{poor sleep quality}=2.128, OR_{unhealthy dietary habits}=2.085, all P<0.05). Compared with elderly patients with chronic diseases without co-morbidity, patients with co-morbidity had higher odds of exacerbation of the original disease/acute readmission and lower odds of new chronic disease (χ²_{primary exacerbation/emergency readmission}=10.726, χ²_{new chronic disease}=5.873 , all P<0.05). Conclusion Gender, age, chronic disease history, BMI, and lifestyle habits are important factors influencing co-morbidity in elderly patients with chronic diseases, and patients with co-morbidity have a relatively poor prognosis.

Objective We aimed to investigate the TCM syndrome characteristics and risk factors of early death in higher-risk patients with myelodysplastic syndromes with excess blasts (HR-MDS-EB). Methods The data of 57 HR-MDS-EB patients in the hematology ward of Xiyuan Hospital, China Academy of Chinese Medical Sciences admitted between January 2013 and August 2020 were analyzed. Patients were divided into an early death group and a non-early death group according to the survival time. The basic information, clinical characteristics, specialized examination, complete blood cell counts and laboratory examination result for first diagnosis, TCM syndrome characteristics, and other data of patients were collected and analyzed to screen the risk factors of death.Results There were statistically significant differences in Charlson comorbidity index (CCI) score, Barthel index score, platelet count at first diagnosis, blast ratio in peripheral blood, lactate dehydrogenase levels, blazing pathogenic heat pattern, and constipation between the early death group and the non-early death group (P<0.05). According to the result of the above univariate analysis, logistic multivariate regression analysis was performed for significant factors P<0.05. CCI score had statistically significant effects on the risk of death (P<0.05).Conclusion Higher CCI score is an independent risk factor for early death in HR-MDS-EB patients.

Compared with conventional vaccines, mRNA vaccines have considerable advantages in design, production, and application, especially in dealing with emerging infectious diseases. Particularly, mRNA vaccines were the first to be recommended by the World Health Organization for emergency use during the COVID-19 pandemic. A key to the design of mRNA vaccines is to ensure the stable and sufficient expression of the encoded protein in the recipient. In recent years, advances have been attained in the experimental and computational research in this area. This review focused on the progress and problems in improving the translation efficiency of mRNA vaccines in recent years, aiming to promote related research.
mRNA Vaccines ; Humans ; Protein Biosynthesis ; Vaccines, Synthetic/immunology* ; COVID-19 Vaccines/immunology* ; COVID-19/prevention & control* ; SARS-CoV-2/genetics* ; RNA, Messenger/genetics*

Objective:To investigate the efficacy and safety of intravenous thrombolysis (IVT) in patients with acute ischemic stroke (AIS) who are currently using direct oral anticoagulants (DOACs).Methods:Patients with AIS admitted to the Stroke Center of Zhengzhou People's Hospital from January 2021 to December 2024 and had taken DOACs within 48 hours prior to onset were included retrospectively. The demographic characteristics, vascular risk factors, laboratory test results, baseline National Institutes of Health Stroke Scale (NIHSS) scores, and other clinical data were collected. The main outcome measure was the functional outcome evaluated using the modified Rankin Scale at 90 days after onset, with a score of 0-2 defined as good outcome. The secondary outcome measures were symptomatic intracranial hemorrhage (sICH) and any bleeding in any location occurring within 36 hours after onset. Multivariate logistic regression analysis was used to determine the independent influencing factors of the outcome. Results:A total of 153 patients were included, with 87 males (56.9%), aged (71.261±6.983) years. Seventy-four patients (48.4%) underwent IVT, and 59 (38.6%) had poor outcome. The good outcome rate in the IVT group was significantly higher than that in the non-IVT group (71.6% vs. 51.9%; χ2=6.274, P=0.012), but there was no significant difference in the incidence of sICH and any bleeding in any location. The baseline NIHSS score of the good outcome group was significantly lower than that of the poor outcome group (8.817±3.677 vs. 11.203±5.060; t=3.361, P<0.001), and the proportion of IVT was significantly higher than that of the poor outcome group (56.4% vs. 35.6%; χ2=6.274, P=0.012). Multivariate logistic regression analysis showed that previous history of stroke or transient ischemic attack (odds ratio [ OR] 3.964, 95% confidence interval [ CI] 1.611-9.753; P=0.003) and high baseline NIHSS score ( OR 1.129, 95% CI 1.034-1.233; P=0.007) were independently associated with the poor outcome, while IVT was independently associated with the good outcome ( OR 0.166, 95% CI 0.068-0.410; P<0.001). Conclusion:For patients with AIS who had taken DOACs within 48 hours before onset, IVT can significantly improve the outcome without increasing the risk of sICH.

Objective:To investigate the risk factors for hemorrhagic transformation (HT) after intravenous thrombolysis (IVT) in patients with acute ischemic stroke (AIS), and the predictive value of Neutrophil to lymphocyte ratio (NLR).Methods:Consecutive patients with AIS received IVT in Zhengzhou People’s Hospital from January 2021 to December 2022 were retrospectively enrolled. HT was defined as no intracranial hemorrhage was found on the first imaging examination after admission, and new intracranial hemorrhage was found on the imaging examination 24 h after IVT or when symptoms worsened. sHT was defined as HT and the National Institutes of Health Stroke Scale (NIHSS) score increased by ≥4 compared to admission or required surgical treatment such as intubation and decompressive craniectomy. The baseline clinical and laboratory data of the patients were collected, and NLR, lymphocyte to monocyte ratio (LMR), and platelet to neutrophil ratio (PNR) were calculated. Multivariate logistic regression analysis was used to identify the independent predictors of HT and sHT, and receiver operating characteristic (ROC) curve was used to analyze the predictive value of NLR for HT and sHT after IVT. Results:A total of 196 patients were included (age 65.37±13.10 years, 124 males [63.3%]). The median baseline NIHSS score was 4 (interquartile range: 2-10). Twenty patients (10.2%) developed HT, and 12 (6.1%) developed sHT. Univariate analysis showed that there were statistically significant differences in age, baseline NIHSS score, creatinine, NLR, and stroke etiology type between the HT group and the non-HT group (all P<0.05); there were statistically significant differences in age, NLR, PNR, creatinine, baseline NIHSS score, and stroke etiological type between the sHT group and the non-sHT group (all P<0.05). Multivariate logistic regression analysis showed that NLR was an independent predictor of HT (odds ratio [ OR] 1.375, 95% confidence interval [ CI] 1.132-1.670; P=0.001) and sHT ( OR 1.647, 95% CI 1.177-2.304; P=0.004) after IVT. The ROC curve analysis showed that the area under the curve for predicting HT by NLR was 0.683 (95% CI 0.533-0.833; P=0.007), the optimal cutoff value was 5.78, the sensitivity and specificity were 55.0% and 84.1%, respectively. The area under the curve for predicting sHT by NLR was 0.784 (95% CI 0.720-0.839; P=0.001), the optimal cutoff value was 5.94, the sensitivity and specificity were 66.67% and 84.24%, respectively. Conclusions:A higher baseline NLR is associated with an increased risk of HT and sHT after IVT in patients with AIS, and can serve as a biomarker for predicting HT and sHT after IVT.

Objective:To investigate the relationship between the overall burden score of cerebral small vessel disease (CSVD) and sudden sensorineural hearing loss (SSNHL) and its severity.Methods:Patients with SSNHL admitted to Zhengzhou People’s Hospital from January 2019 to June 2022 were used as the case group, and age- and gender-matched patients with tension headache or benign positional vertigo were used as the control group. MRI was used to evaluate the phenotype of CSVD, including white matter hyperintensities (WMHs), cerebral microbleeds (CMBs), enlarged perivascular spaces (EPVS), and lacune of presumed vascular origin, and then the total burden score of CSVD was calculated. Multivariate logistic regression analysis was used to determine the independent factors of SSNHL. Ordinal logistic regression analysis was used to investigate the relationship between the overall burden of CSVD and the severity of hearing loss in patients with SSNHL. Results:A total of 86 patients with SSNHL and 90 age- and gender-matched controls were included. There were statistically significant differences in triglycerides, high-density lipoprotein cholesterol, fasting blood glucose levels, WMHs overall score classification, periventricular WMHs score classification, deep WMHs score classification, CMBs, moderate to severe EPVS, CSVD overall burden score and classification between the two groups (all P<0.05). Multivariate logistic regression analysis showed that after adjusting for high-density lipoprotein cholesterol, fasting blood glucose ≥6.1 mmol/L (odds ratio [ OR] 2.149, 95% confidence interval [ CI] 1.042-4.432; P=0.038), triglycerides ≥1.7 mmol/L ( OR 7.012, 95% CI 2.962-16.597; P=0.000), CSVD overall burden score >1 (compared to 0 point, 2 points: OR 4.095, 95% CI 1.364-12.291, P=0.012; 3 points: OR 5.776, 95% CI 1.888-17.614, P=0.002; 4 points: OR 7.900, 95% CI 2.526-24.706; P=0.000) were significantly independently correlated with SSNHL. Ordinal logistic regression analysis showed that after adjusting for age and fasting blood glucose, the flat decline type hearing loss ( OR 1.276, 95% CI 1.131-1.618; P=0.044) and total deafness type hearing loss ( OR 1.029, 95% CI 1.002-1.058; P=0.038), and the overall burden of CSVD being moderate ( OR 1.318, 95% CI 1.036-1.677; P=0.025) and severe ( OR 2.330, 95% CI 1.232-4.406; P=0.009) were significantly independent associated with the degree of hearing loss in patients with SSNHL. Conclusion:The CSVD total burden score of moderate to severe degree is independently associated with SSNHL and the degree of hearing loss.