Reactive oxygen species (ROS) are major contributors to radiation therapy-induced side effects in cancer patients. A fusion antioxidant enzyme comprising glutathione S-transferase (GST), superoxide dismutase 1 (SOD1), and a transmembrane peptide has been shown to effectively mitigate ROS-induced damage. To enhance its targeting capability, the fusion protein was further modified by incorporating a matrix metalloproteinase-2/9 substrate peptide (X) and the transmembrane peptide R9, yielding the antioxidant enzyme GST-SOD1-X-R9 (GS1XR). This modification reduced its transmembrane ability in tumor cells, thereby selectively protecting normal cells from oxidative stress. However, the use of non-human GST poses potential immunogenicity risks. In this study, we employed seamless cloning technology to construct an expression vector containing the human GST gene to replace the non-human GST gene, and then expressed and purified novel fusion antioxidant enzymes GS1R and GS1XR. The protective effects of newly constructed GS1R and GS1XR against hydrogen peroxide (H₂O₂)-induced oxidative damage in L-02 cells were then evaluated using GS1 as a control. Enzymatic activity assays revealed that the specific activity of GST in GS1XR remained unchanged compared to the unmodified protein, while SOD activity was enhanced. Exposure to 200 μmol/L H₂O₂ transiently activated the nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway; however, this activation diminished after 24 h, reducing cell viability to 48.4%. Both GS1R and GS1XR effectively scavenged intracellular ROS, directly counteracting oxidative stress and promoting Nrf2 nuclear translocation, thereby activating antioxidant pathways and restoring cell viability to normal levels. The two enzymes showed comparable efficacy. In contrast, GS1, lacking transmembrane capability, was restricted to scavenging extracellular ROS and provided only limited protection. In conclusion, both novel fusion antioxidant enzymes demonstrated significant potential in safeguarding normal cells from ROS-mediated oxidative damage. The findings provide a foundation for further investigation in related field.
Humans ; Oxidative Stress/drug effects* ; Hydrogen Peroxide ; Antioxidants/metabolism* ; Glutathione Transferase/metabolism* ; Recombinant Fusion Proteins/pharmacology* ; Superoxide Dismutase-1 ; Reactive Oxygen Species/metabolism* ; Superoxide Dismutase/biosynthesis*

The study aims to investigate the impacts of prolyl oligopeptidase (POP) on the replication of foot-and-mouth disease virus (FMDV) in BHK-21 cells. Firstly, the effects of FMDV replication on POP expression in BHK-21 cells were analyzed by Western blotting and Real-time reverse transcription polymerase chain reaction (RT-qPCR). Secondly, a eukaryotic expression plasmid for POP was constructed, and the effects of POP overexpression on the replication of two different serotypes of FMDV were assessed by Western blotting, RT-qPCR, and virus titer assays. Thirdly, specific small interfering RNAs (siRNAs) targeting POP were synthesized, and their efficiency in interfering with endogenous POP expression was identified by RT-qPCR. The impacts of downregulating endogenous POP expression on FMDV replication were further evaluated by Western blotting, RT-qPCR, and virus titer assays. The results indicated that FMDV infection did not significantly affect POP expression in BHK-21 cells. Overexpression of POP dose-dependently enhanced the replication of both FMDV/O and FMDV/A serotypes. Conversely, siRNA-mediated downregulation of endogenous POP expression markedly suppressed FMDV/O replication. This study is the first to demonstrated that the role of the host POP protein in promoting FMDV replication in BHK-21 cells, thereby providing a critical theoretical foundation and potential molecular targets for developing efficient candidate cell strains for foot-and-mouth disease inactivated vaccines.
Foot-and-Mouth Disease Virus/genetics* ; Virus Replication/genetics* ; Prolyl Oligopeptidases ; Serine Endopeptidases/physiology* ; Animals ; Cell Line ; RNA, Small Interfering/genetics* ; Foot-and-Mouth Disease/virology* ; Cricetinae

Objective To compare the analgesic effects and adverse reactions between intercostal nerve block (ICNB) and patient controlled intravenous analgesia (PCIA). Methods From August 2022 to January 2023, 180 patients with lung cancer who underwent thoracoscopic surgery were randomly divided into two groups: ICNB group (n=90) and PCIA group (n=90). The postoperative pain degree (VAS), location, nature; adverse events, such as nausea, vomiting, and dizziness; and other clinical symptoms were analyzed. Results The most common site of postoperative pain in both groups was surgical incision, and the nature of pain was distending pain. At 12 and 24 h after the operation, the pain degree in the ICNB group (1.10±0.91, 3.12±1.29) was markedly lower than that in PCIA group (1.44±0.86, 4.32±1.30, P=0.010, P<0.001). The incidence of nausea, vomiting, and dizziness in the ICNB group (5.56%, 23.33%) was noticeably lower than that in the PCIA group (35.56%, 51.11%, P<0.001, P<0.001). Total hospitalization expense in the ICNB group (41 043.16±10 885.63 yuan) was significantly lower than that in PCIA group (45 283.99±11 036.36 yuan, P=0.010). Conclusion The analgesic effect of intercostal nerve block is better than that of patient-controlled intravenous analgesia pump in patients with lung cancer after minimally invasive surgery, and the incidence of adverse reactions is low.

Objective: To investigate the creation of smoke-free environments in public places in Hangzhou City, so as to provide insights into effective implementation of the tobacco control policy. Methods: The party and government administrations at each level, medical institutions, educational places, restaurants and entertainment places, and open public places were enrolled. The creation of smoke-free environments was investigated in these places through undercover investigation with field observations and concealed photography by a third-party professional investigation company from November to December, 2022. The building of smoke-free environments (totally 60 scores) and no smoking indoors (totally 40 scores) were evaluated according to the Criteria for Scoring of Smoke-free Organizations in Hangzhou City. Results: Totally 909 places were investigated, and the comprehensive score of smoke-free environment building was (82.83±14.13) points. There were 285 party and government administrations with a comprehensive score of (84.19±12.85) points, 65 medical institutions with a comprehensive score of (90.35±6.95) points, 65 educational places with a comprehensive score of (83.43±16.81) points, 403 dining and entertainment places with a comprehensive score of (80.68±14.75) points, and 91 open public places, with a comprehensive score of (82.34±14.77) points. There were 397 places with standardized tobacco control tips at entrances (43.67%), 308 places with tobacco control signs posted as required (33.88%), 707 places that set outdoor smoking areas correctly (77.78%), 68 places with smoking paraphernalia (7.48%), 28 places with tobacco sales (3.08%). There were 732 places without signs of indoor smoking (80.53%), 850 places without indoor smoking (93.51%) and 24 places without dissuading from smoking (2.64%). Conclusion The indoor no-smoking is overall satisfactory in public places in Hangzhou City; however, standardizing no-smoking tips at entrances, standardizing the posting of no-smoking signs and assignment of tobacco control materials remain to be improved.

ObjectiveTo investigate the effects and molecular mechanism of ursolic acid on the proliferation and apoptosis of colorectal cancer cells. MethodThe proliferation inhibition rate of human colorectal cancer RKO cells treated with different concentrations of ursolic acid （0， 5， 10， 15， 20， 25， 30 μmol·L^-1） was detected by cell counting kit-8 （CCK-8）， and the half maximal inhibitory concentration （IC₅₀） at 24 h and 48 h was calculated. According to the IC₅₀ of RKO cells treated with ursolic acid for 24 h， two concentrations were selected for subsequent experiments. The colony formation assay was used to detect the proliferation ability of the cells and flow cytometry was used to detect the apoptosis rate and cell cycle arrest after treatment of RKO cells with ursolic acid. After treatment of RKO cells with ursolic acid for 24 hours， the expression of B-cell lymphoma 2 （Bcl-2）-associated X protein （Bax） in RKO cells， Bcl-2 in Raji cells， PMA responsive gene in T lymphocyte （Noxa）， cyclin-dependent kinase inhibitor 1A （p21）， cyclin-dependent kinase inhibitor 1B （p27）， cyclin-dependent kinase 4 （CDK4）， protein kinase B （Akt）， phosphorylated Akt （p-Akt）， forkhead transcription factor O3a （FoxO3a）， and phosphorylated FoxO3a （p-FoxO3a） was determined by Western blot. ResultCompared with the blank group， the ursolic acid groups could inhibit the viability of RKO cells （P<0.05， P<0.01）， and the colony formation rates of RKO cells in the ursolic acid groups were reduced （P<0.05， P<0.01） in a concentration-dependent manner. The cells in the ursolic acid group （20 μmol·L^-1） experienced cell cycle arrest， which increased in the early stage of synthesis， ie， the G₀/G₁ phase （P<0.05） as compared with the results in the blank group. Compared with the blank group， the ursolic acid groups （15 and 20 μmol·L^-1） showed increased protein expression of p21 and p27， decreased expression of CDK4 protein （P<0.05， P<0.01）， and increased apoptosis rate， and the ursolic acid group （20 μmol·L^-1） showed increased protein expression of Bax and Noxa and decreased expression of Bcl-2 （P<0.05， P<0.01）. In terms of mechanism， compared with the blank group， the ursolic acid group （20 μmol·L^-1） down-regulated the expression of p-Akt protein and up-regulated the expression of p-FoxO3a （P<0.05， P<0.01）， and there was no significant change in the total protein of Akt and FoxO3a. ConclusionUrsolic acid can effectively inhibit the proliferation of colorectal cancer RKO cells and promote cell apoptosis， which may be related to the Akt/FoxO pathway.

Progressive functional deterioration in the cochlea is associated with age-related hearing loss (ARHL). However, the cellular and molecular basis underlying cochlear aging remains largely unknown. Here, we established a dynamic single-cell transcriptomic landscape of mouse cochlear aging, in which we characterized aging-associated transcriptomic changes in 27 different cochlear cell types across five different time points. Overall, our analysis pinpoints loss of proteostasis and elevated apoptosis as the hallmark features of cochlear aging, highlights unexpected age-related transcriptional fluctuations in intermediate cells localized in the stria vascularis (SV) and demonstrates that upregulation of endoplasmic reticulum (ER) chaperon protein HSP90AA1 mitigates ER stress-induced damages associated with aging. Our work suggests that targeting unfolded protein response pathways may help alleviate aging-related SV atrophy and hence delay the progression of ARHL.
Mice ; Animals ; Transcriptome ; Aging/metabolism* ; Cochlea ; Stria Vascularis ; Presbycusis

Objective:To discuss the long-term survival and risk factors of thyroid cancer in the real world in China.Methods:The clinical data of thyroid cancer patients who underwent initial surgery from Apr. 1998 to Dec. 2018 were retrospectively analyzed, including patients’sex, age, surgical records, pathology, hospitalization records and follow-up. According to the prognosis, the patients were divided into disease-free survival group and recurrence/metastasis/death group. Univariate analysis and multivariate regression analysis were conducted to analyze the risk factors affecting the prognosis of thyroid cancer. The clinical features and prognostic risk factors of thyroid cancer patients were investigated.Results:A total of 2038 cases were collected, and the longest follow-up time was more than 20 years. A total of 1876 cases were included in the study, 162 cases were lost, and the rate of follow-up was 7.9%. Among them, 1858 survived, the overall survival rate was 99.04%; 18 died, and the overall mortality rate was 0.96%. According to the prognosis of thyroid cancer, the patients were divided into 2 groups, including 1808 cases in the disease-free survival group and 68 cases in the relapsed-metastatic-death group. The study found that there were statistical differences between the two groups in terms of patients’age [ (45.40±11.016) vs (51.53±15.199, P=0.000) , the male ratio (32.854%, 48.529%, P=0.001) , whether tumor breaks through capsule (20.077%, 33.823%, P=0.006) , central lymph node metastasis (48.834%, 70.588%, P=0.001) and lateral lymph node metastasis (31.084%, 55.882%, P=0.000) , and there was no difference between the number of tumor lesions. Conclusion:Thyroid cancer has a good prognosis, but according to the characteristics of patients with thyroid cancer in my country, it should still be treated early in the clinic, and the standardization and thoroughness of surgery should be adhered to during the treatment.

Objective:To investigate the methylation status of SDC2, PPP2R5C and ADHFE1 genes in stool and their values in the screening of colorectal cancer and precancerous lesions.Methods:From August 2020 to March 2021, 64 patients with colorectal cancer, 72 patients with adenoma, 33 patients with hyperplastic polyps and 59 healthy people were recruited from Qingdao Central Hospital Affiliated to Qingdao University, and the morning stool samples were collected from the research subjects. The genomic DNA was extracted and modified with sulfite. The methylation status of SDC2, PPP2R5C and ADHFE1 genes were detected by methylation specific polymerase chain reaction (MSP), and the fecal occult blood test (FOBT) was performed. Taking the pathological results as the gold standard, receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to compare the effect of combined detection of methylation of three genes and FOBT in predicting colorectal cancer and precancerous lesions. R-Studio software was used to construct a nomogram for the prediction of colorectal cancer with combined detection of gene methylation in stool and other clinical features, and the calibration and validation were performed.Results:The positive rates of combined detection of methylation of SDC2, PPP2R5C and ADHFE1 genes in stool were higher than those of FOBT in colorectal cancer+adenoma [74.3% (101/136) vs. 47.1% (64/136), χ2 = 23.20, P = 0.001], colorectal cancer [90.6% (58/64) vs. 70.3% (45/64), χ2 = 8.91, P = 0.003] and adenoma [59.7% (43/72) vs. 26.4% (19/72), χ2 = 14.43, P = 0.002]. There was no significant difference in the positive rates in hyperplastic polyps [21.2% (7/33) vs. 6.1% (2/33), χ2 = 0.12, P = 0.125] and healthy controls [10.2% (6/59) vs. 8.5% (5/59), χ2 = 4.01, P = 1.000]. The combined detection of gene methylation was better than FOBT in the prediction of colorectal cancer + adenoma [AUC: 0.85 (95% CI 0.80-0.91) vs. 0.71 (95% CI 0.64-0.78), P < 0.05], especially in the prediction of adenoma [AUC: 0.82 (95% CI 0.74-0.89) vs 0.64 (95% CI 0.57-0.69), P < 0.001]. The sensitivity and specificity of ADHFE1 gene methylation status in predicting colorectal cancer were high (90.6% and 96.6%). In colorectal cancer patients over 50 years old, the positive rate of combined detection of gene methylation was higher than that of FOBT [90.2% (55/61) vs. 68.9% (42/61), P < 0.05]. The nomogram calibration curve for predicting colorectal cancer constructed based on the combined detection of gene methylation and each clinical feature showed a high degree of concordance between the predicted and observed diagnostic performance of colorectal cancer. Conclusions:The methylation levels of SDC2, PPP2R5C AND ADHFE1 genes in stool are increased in patients with colorectal cancer or adenoma. The combined detection of gene methylation is expected to be a non-invasive method for the screening of colorectal cancer and precancerous lesions.

Objective To study the effects of integrated orthopedic rehabilitation pathway on motor function in six months after total knee arthroplasty (TKA), including pain, stiffness, range of motion and muscle strength, etc. Methods From March, 2016 to March, 2019, 180 patients who underwent TKA and treated with integrated orthopedic rehabilitation pathway were enrolled. Age, gender, operation time, time of follow-up, the scores of Hospital for Special Surgery-Knee Scale (HSS-KS) and Western Ontario and McMaster Universities osteoarthritis index (WOMAC) at preoperative/postoperative/one-month after operation/three-month after operation/six-month after operation time points were collected. The sub items, such as muscle strength, range of motion, flexion deformity, pain, stiffness, functional difficulty were primarily focused on. Results A total of 42 patients were followed up for three months and 22 patients were followed up for six months. There was no significant difference in the scores of HSS-KS and WOMAC before and after operation (P > 0.05). Within three months after operation, the HSS-KS scores gradually increased (P < 0.05) and the WOMAC scores gradually decreased (P < 0.05). The active knee flexion range of motion and knee extensor muscle strength scores of HSS-KS significantly decreased after operation (P < 0.05), and gradually recovered one month and three months after operation (P < 0.05). The flexion deformity scores of HSS-KS increased after operation (P < 0.05), decreased one month after operation (P < 0.05), and got a trend of incensement again three months after operation. The pain score of WOMAC decreased continuously within three months after operation (P < 0.05); the stiffness score of WOMAC did not change after operation (P > 0.05), decreased significantly one month after operation (P < 0.05), and did not change three months after operation (P > 0.05). The degree of functional difficulty of WOMAC decreased after operation (P < 0.05), and improved continuously within six months after operation (P < 0.05). Conclusion The overall function after TKA shows a trend of improvement within three months, and there is no obvious improvement from three to six months after operation. The flexion deformity score showed a downward trend in one month after operation, and it could be improved again after strengthening rehabilitation, which needs more attention in the postoperative rehabilitation.

Animals ; Mice ; Motor Neurons/physiology* ; Spinal Cord