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*

OBJECTIVE To explore the effects of 5，10-methylenetetetrahydrofolate reductase （MTHFR） gene polymorphism on the adverse reactions in patients with osteosarcoma after the first high-dose methotrexate （HD-MTX） treatment. METHODS A prospective study was conducted to include 53 patients with osteosarcoma treated with HD-MTX at the first admission in General Hospital of Eastern Theater Command. The dose of MTX was evaluated according to the polymorphism of rs1801133 in the METHFR gene and demographic factors， then whole pharmaceutical monitoring was conducted. The data on liver toxicity， renal toxicity， hematological toxicity， and gastrointestinal reaction were collected after the first chemotherapy cycle. Single factor analysis and binary Logistic regression analysis were used to analyze the correlation between MTX dose， 24 h blood drug concentration， and rs1801133 locus genotype with four adverse reactions. RESULTS The MTX dosage in patients with CC wild type was significantly higher than that in TT mutant type （7.97 g/m2 vs. 6.98 g/m2， P＝0.030）， but this difference did not affect the 0 h and 24 h blood drug concentrations of MTX. The above four adverse reactions were not related to the dose of MTX. The results of binary Logistic regression analysis showed that carrying one T allele increased the risk of developing hematological toxicity by 4.13 times（95% confidence interval：1.35-12.62，P＝0.013）. When 24 h plasma concentration threshold of MTX was set to 2.65 µmol/L， the sensitivity and specificity of predicting liver function damage were 53.33% and 86.96%， respectively； when the threshold was set to 7.28 μmol/L， the sensitivity and specificity of predicting renal damage were 100% and 81.63%. CONCLUSIONS The polymorphism of the rs1801133 in the MTHFR gene is associated with hematological toxicity of MTX. Patients who take HD-MTX for the first time and carry the T allele have a high risk of hematological toxicity. The 24 h plasma concentration of MTX is related to liver toxicity and renal toxicity. In addition， monitoring the 24 h blood drug concentration can predict liver and renal toxicity， and take early intervention.

Intestinal dysbiosis and disrupted bile acid(BA)homeostasis are associated with obesity,but the precise mechanisms remain insufficiently explored.Hepatic protein phosphatase 1 regulatory subunit 3G(PPP1R3G)plays a pivotal role in regulating glycolipid metabolism;nevertheless,its obesity-combatting potency remains unclear.In this study,a substantial reduction was observed in serum PPP1R3G levels in high-body mass index(BMI)and high-fat diet(HFD)-exposed mice,establishing a positive correlation between PPP1R3G and non-12α-hydroxylated(non-12-OH)BA content.Additionally,hepatocyte-specific overexpression of Ppp1r3g(PPP1R3G HOE)mitigated HFD-induced obesity as evidenced by reduced weight,fat mass,and an improved serum lipid profile;hepatic steatosis alleviation was confirmed by normalized liver enzymes and histology.PPP1R3G HOE considerably impacted systemic BA homeostasis,which notably increased the non-12-OH BAs ratio,particularly lithocholic acid(LCA).16S ribosomal DNA(16S rDNA)sequencing assay indicated that PPP1R3G HOE reversed HFD-induced gut dysbiosis by reducing the Firmicutes/Bacteroidetes ratio and Lactobacillus population,and elevating the relative abundance of Blautia,which exhibited a positive correlation with serum LCA levels.A fecal microbiome transplantation test confirmed that the anti-obesity effect of hepatic PPP1R3G was gut microbiota-dependent.Mechanistically,PPP1R3G HOE markedly suppressed hepatic cholesterol 7α-hydroxylase(CYP7A1)and sterol-12α-hydroxylase(CYP8B1),and concurrently upregulated oxysterol 7-α hydroxylase and Takeda G protein-coupled BA receptor 5(TGR5)expression under HFD conditions.Furthermore,LCA administration significantly mitigated the HFD-induced obesity phenotype and elevated non-12-OH BA levels.These findings emphasize the significance of hepatic PPP1R3G in ameliorating diet-induced adiposity and hepatic steatosis through the gut microbiota-BA axis,which may serve as potential ther-apeutic targets for obesity-related disorders.

Insulin-like growth factor-1 receptor (IGF-1R) has been made an attractive anticancer target due to its overexpression in cancers. However, targeting it has often produced the disappointing results as the role played by cross talk with numerous downstream signalings. Here, we report a disobliging IGF-1R signaling which promotes growth of cancer through triggering the E3 ubiquitin ligase MEX3A-mediated degradation of RIG-I. The active β-arrestin-2 scaffolds this disobliging signaling to talk with MEX3A. In response to ligands, IGF-1Rβ activated the basal βarr2 into its active state by phosphorylating the interdomain domain on Tyr64 and Tyr250, opening the middle loop (Leu130‒Cys141) to the RING domain of MEX3A through the conformational changes of βarr2. The models of βarr2/IGF-1Rβ and βarr2/MEX3A could interpret the mechanism of the activated-IGF-1R in triggering degradation of RIG-I. The assay of the mutants βarr2^Y64A and βarr2^Y250A further confirmed the role of these two Tyr residues of the interlobe in mediating the talk between IGF-1Rβ and the RING domain of MEX3A. The truncated-βarr2 and the peptide ATQAIRIF, which mimicked the RING domain of MEX3A could prevent the formation of βarr2/IGF-1Rβ and βarr2/MEX3A complexes, thus blocking the IGF-1R-triggered RIG-I degradation. Degradation of RIG-I resulted in the suppression of the IFN-I-associated immune cells in the TME due to the blockade of the RIG-I-MAVS-IFN-I pathway. Poly(I:C) could reverse anti-PD-L1 insensitivity by recovery of RIG-I. In summary, we revealed a disobliging IGF-1R signaling by which IGF-1Rβ promoted cancer growth through triggering the MEX3A-mediated degradation of RIG-I.

Objective:To investigate the expression level of circular RNA circ_0020123 in ovarian cancer (OC) , and then to evaluate the diagnostic value of circ_0020123 level in ovarian cancer.Methods:Fifty-five ovarian cancer patients who were diagnosed and treated in Hangzhou Linping District Maternal and Child Health Hospital from Oct. 2016 to Mar. 2017 were selected, and serum was collected for high-throughput gene expression sequencing analysis. The circ_0020123 level in serum of ovarian cancer patients and healthy controls was determined by real-time quantitative PCR, and the diagnostic efficacy of circ_0020123 level on ovarian cancer was evaluated by drawing receiver operating characteristic curve (ROC) . The correlation between the expression of circ_0020123 and the clinical characteristics of ovarian cancer patients was investigated by t-test. The survival curve was drawn to analyze the relationship between the serum circ_0020123 level and the prognosis and survival of ovarian cancer patients. Western blot was used to detect the expression levels of autophagy-related proteins LC3-II/LC3-I and p62.Results:Compared with the serum of healthy people (1±0.25) , the expression of circ_0020123 was significantly up-regulated in the serum of ovarian cancer patients (1.24±0.23) ( t=5.23, P<0.001) . The ROC curve showed that circ_0020123 had high diagnostic performance for the detection of ovarian cancer specimens (AUC=0.7579, P<0.001) . Comprehensive analysis of t test and logistic analysis showed that the high expression of circ_0020123 was significantly correlated with FIGO stage ( t=5.46, P<0.001) and tumor size ( t=6.37, P<0.001) . The measurement of survival data showed that the 5-year overall survival of the circ_0020123 high expression group was significantly shorter than that of the circ_0020123 low expression group ( P=0.019) . Knockdown of circ_0020123 in cells down-regulated the expression of autophagy-related proteins LC3-II/LC3-I (0.45±0.05) ( t=9.31, P=0.001) , and up-regulated the expression of p62 (1.94±0.11) ( t=12.62, P<0.001) . Conclusion:The results of this study indicate that circ_0020123 is an important ovarian cancer-related circular RNA, which provides a potential diagnostic, prognostic biomarker and therapeutic target for ovarian cancer patients.

Objective:To understand the status quo and problems of clinical nursing fund paper publication in China from 2015 to 2018.Methods:The clinical nursing fund papers published in 12 core nursing journals and published by Chinese Authors in WOS database were analyzed by using bibliometrics methodology. Information analyzed including year distribution, number of authors, regional distribution, the number of institutions, research nature, research type, statistical application, specialty distribution and research content.Results:From 2015 to 2018, the proportion of clinical nursing fund papers was increasing, the proportion of funded papers in Chinese and WOS was 44.06% and 51.46% respectively, the main funding sources of each project came from provinces or direct-controlled municipalities level(42.02% in Chinese and 33.06% in WOS). Chinese fund papers were mainly co-authored by four people (20.18%) and mostly from the same institution (67.83%), the fund papers collected by WOS were mainly co-authored by six people (21.14%) and mainly produced by two institutions (40.65%). Clinical trials (26.64%) were the main research types of Chinese funded papers, and statistical inference (73.15%) was the main application of statistical methods. Jiangsu (14.58%) produced the most Chinese fund papers, and the first authors of the fund papers collected by WOS mostly came from Taiwan (43.90%); the main research topics were oncology nursing (16.65% in Chinese and 22.49% in WOS) and disease and symptom nursing (23.57% in Chinese and 23.04% in WOS).Conclusions:The number of papers on clinical nursing fund was increasing year by year, the application of statistics is basically stable, the sources of fund funding were increasingly diversified, and the trend of scientific research cooperation was increasing. Economically developed areas were the main force of scientific research output. The focus of clinical nursing research was oncology nursing and disease and symptom nursing.

Objective: To evaluate the feasibility of expanded indication for endoscopic submucosal dissection (ESD) in undifferentiated early gastric cancer, to investigate the risk factors of lymph node metastasis (LNM), so as to provide the theoretical evidence for the choice of treatment. Methods: From June 2007 to December 2018, at the Affiliated Hospital of Qingdao University, the clinical and pathological data of 807 patients with undifferentiated early gastric cancer and undergoing gastrectomy plus lymphadenectomy were retrospectively analyzed. Chi-square test was performed to analyze the correlation between clinicopathologic characteristics of early gastric cancer and LNM. Multivariate logistic regression model was used to analyze the independent risk factor of LNM. Results: LNM was found in 17.2% (139/807) patients with undifferentiated early gastric cancer. And no LNM was detected in 110 patients who met the expanded indication of ESD. The results of univariate analysis indicated that LNM was significantly associated with increased carcinoembryonic antigen (CEA), tumour size, gross type, ulcer, invasion depth, lymphovascular invasion and perineural invasion (χ²=4.500, 13.332, 16.611, 6.083, 51.064, 0.564 and 17.006, all P<0.05). The results of multivariate analysis demonstrated that the maximum diameter of tumor over 20 mm (odds ratio (OR)=1.606, 95% confidence interval (CI) 1.021 to 2.526, P=0.040), lymphovascular invasion (OR=16.835, 95%CI 10.510 to 26.966, P<0.01), the depth of submucosal superficial invasion (≤500 μm ; OR=1.962, 95%CI 1.022 to 3.765, P=0.043) and the depth of submucosal deep invasion (>500 μm ; OR=3.014, 95%CI 1.753 to 5.181, P<0.01) were independent risk factors of LNM in early gastric cancer. Conclusions The expanded ESD indication of undifferentiated early gastric cancer is applicable for endoscopic treatment considering the low risk of LNM. In early undifferentiated gastric cancer, maximum diameter of tumor over 20 mm, lymphovascular invasion, submucosal superficial and deep invasion are the independent risk factors of LNM.

Sinus floor elevation was needed in 11 patients having 15 implant sites with the residual bone height (RBH) was less than 10 mm in the posterior maxillary region from Feb to May 2017. The RBH ranged from 3.10 to 8.34 mm [averaged (6.18±1.60) mm]. RBH<6 mm was observed in 40% implant sites (6 implant sites) and RBH≥6 mm was observed in 60% implant sites (9 implant sites). The thickness of the sinus floor membrane correspond to the implant site measured by cone beam CT (CBCT) ranged from 0.50 to 4.24 mm [averaged (1.21±0.92) mm]. Sequential drills with stops were used to perforate the cortical bone of the sinus floor firstly, then the transcrestal around detached sinus floor elevation technique (TADSFET) was carried with osteotomes. Anorganic bovine bone was used as the augmentation material.Fifteen implants were placed in 15 implant sites. CBCT pictures showed that there was a smooth and continuous tent-shaped apophysis on each lifted site and no air fluid level was observed in the sinus immediately after operation. The mean elevated height of the 15 implant sites was (7.83±1.57) mm (ranged from 5.94 to 11.01 mm). The mean follow-up time was 7.91 months (7-10 months). The survival rate was 100% during the follow up period.

There are significant individual differences in the antiplatelet effects of aspirin.Three single nucleotide polymorphisms (SNPs),rs5918,rs12041331 and rs730012,are reported to significantly correlate with the efficacy and side effects of aspirin.In the present study,the genotyping method of the three SNPs was established based on the combination of polymerase chain reaction and pyrosequencing technology.Amplification and sequencing primers were designed independently;the amplification conditions were optimized to amplify the three SNPs in the same condition.The sensitivity of the method was detected using original genome DNA at different concentrations.In order to testify the accuracy of the method,the proposed method and Sanger sequencing technology were both used to genotype the three SNPs in 20 blood samples.The results demonstrated that the genotyping method of aspirin-related SNPs was successfully established,with the detection limit being as low as 0.4 ng genome DNA.The genotype results of 20 samples by the proposed method were exactly the same as that of Sanger sequencing.It is evident that the proposed method is sensitive and accurate.

Objective To investigate the acceptability and related factors of an "on-demand"pre-exposure prophylaxis (PrEP) to prevent HIV transmission among MSM in Shenyang.Methods MSM recruited by non-probability sampling method and questionnaire survey conducted by investigators to collect information on social and behavioral characteristics,awareness of PrEP,Truvada and the acceptability of two different PrEP-based trials.Multivariate logistic regression was employed for statistical analysis.Results Among the 292 respondents,34.2％ had heard of PrEP and 58.2％ (170/292) reported were interested in participating a PrEP trial-"on-demand" use or 48.3％ (141/292) interested on "daily" use (x2=5.785,P=0.02).Factors independently associated with those "on-demand" would include:having more than 2 male sexual partners during the past 6 month (aOR=1.7,95％CI:1.1-2.7),concerning on the positive effect of PrEP (vs.side effects) (aOR=6.4,95％CI:2.2-18.9),having an HIV-infected sexual partners (aOR=8.1,95％ CI:1.0-63.3) and self-reported high risk for HIV (aOR=2.6,95％CI:1.2-6.0).The last two factors were only associated with the "on-demand" group.Conclusions "On-demand" PrEP (as opposed to daily) seemed a more feasible prevention strategy on HIV and particularly in those having high risk behavior of HIV.For those who could not follow the daily medication or having HIV risk perception,"On-demand" basis PrEP trial should be recommended for them to follow.