Engineering stable and non-immunogenic immunoenzymes for cancer therapy via in situ generated prodrugs.

Engineering human enzymes for therapeutic applications is attractive but introducing new amino acids may adversely affect enzyme stability and immunogenicity. Here we used a mammalian membrane-tethered screening system (ECSTASY) to evolve human lysosomal beta-glucuronidase (hBG) to hydrolyze a glucuronide metabolite (SN-38G) of the anticancer drug irinotecan (CPT-11). Three human beta-glucuronidase variants (hBG3, hBG10 and hBG19) with 3, 10 and 19 amino acid substitutions were identified that display up to 40-fold enhanced enzymatic activity, higher stability than E. coli beta-glucuronidase in human serum, and similar pharmacokinetics in mice as wild-type hBG. The hBG variants were two to three orders of magnitude less immunogenic than E. coli beta-glucuronidase in hBG transgenic mice. Intravenous administration of an immunoenzyme (hcc49-hBG10) targeting a sialyl-Tn tumor-associated antigen to mice bearing human colon xenografts significantly enhanced the anticancer activity of CPT-11 as measured by tumor suppression and mouse survival. Our results suggest that genetically-modified human enzymes represent a good alternative to microbially-derived enzymes for therapeutic applications.

Suggestions on Relieving Physical Anxiety of Medical Workers and Improving Physical and Mental Health Under the COVID-19 Epidemic-A Case Study of Meizhou City.

The study examined the effects of swimming pools on healthcare professionals' willingness to engage in recreational activities, physical anxiety, and physical and mental well-being in the context of COVID-19. The research adopted the mixed research method, used SPSS 26.0 statistical software to test the reliability of the questionnaire, and then collected 840 valid questionnaires; first analyzed the data with basic statistics, t-test, ANOVA, and PPMCC test methods, and then used the interview method to collect expert opinions. A multi-check approach assembled all data and discussions. The study found that the use of personnel dynamic tracking systems or measures, combined with sodium hypochlorite and repeated filtration to stabilize water quality, could maintain the confidence of most medical workers in the swimming pool sports environment for epidemic prevention and avoid violations. The government could formulate safety prevention and control mechanisms in traffic and establish appropriate traffic routes. Next, formulated a prescription for swimming or other physical activity mechanisms for men aged 31-50 and redesigned measures for medical staff over 51 years old to have tense head issues and physical fatigue, promote blood circulation and improve sleep quality. This will promote the purpose of relieving stress and regulating the physical and mental health of medical staff after engaging in swimming.

Effects of Sports Massage on the Physiological and Mental Health of College Students Participating in a 7-Week Intermittent Exercises Program.

The purpose of the research is to analyze the improvement in the physical and mental health of college students after intermittent exercises are performed by massage. The present study employed a mixed research method. An experimental study was conducted to analyze the current status of the volunteers' sports performance and body composition, and then a questionnaire was designed for the subjects' physical and mental health. The data were then analyzed using SPSS 26.0 software for statistical analysis such as t-test and ANOVA. The subjects were then interviewed to collect their opinions on the study results, and finally, the results were explored by multivariate analysis. The study found that intermittent exercise can help university students develop physical fitness and performance, improve body composition, and regulate physical and mental health. The combination of intermittent exercise with sports massages further enhanced the performance of sit-ups and standing long jump, improve blood pressure, BMI, and self-confidence, as well as reducing suicidal tendencies (experimental group > control group). However, intermittent exercise participants still experienced fatigue, headache, emotional loss, and fear of depression, and the addition of sports massage did not significantly improve flexibility and cardiorespiratory endurance (control group > experimental group).

Ergostatrien-3ß-ol (EK100) from Antrodia camphorata Attenuates Oxidative Stress, Inflammation, and Liver Injury In Vitro and In Vivo.

Hepatic ischemia/reperfusion (IR) injury is a complication that occurs during liver surgery, whereby hepatic tissue is injured by oxygen deficiency during ischemia, then further damaged by a cascade of inflammatory and oxidative insults when blood is resupplied during reperfusion. Antrodia camphorata is an indigenous fungus in Taiwan and an esteemed Chinese herbal medicine with various bioactivities. This study examined the effect of ergostatrien-3ß-ol (EK100), an active compound found in both the fruiting body and mycelia of A. camphorata, on IR injury pathologies in rats and cell models of oxidative and inflammatory stress. Male Sprague-Dawley rats were randomly assigned to receive a vehicle or 5 mg/kg EK100 prior to hepatic IR injury induced by 1 h ischemia followed by 24 h reperfusion, or a sham operation. RAW 264.7 murine macrophages and HepG2 hepatocytes were pretreated with EK100, then inflammation was induced with lipopolysaccharides in the former and oxidative stress was induced with hydrogen peroxide in the latter. EK100 decreased IR-induced elevation in serum levels of alanine aminotransferase and aspartate aminotransferase and lowered levels of the inflammatory cytokines tumor necrosis factor-α, interleukin (IL)-6, and IL-1ß. In addition, EK100 significantly reduced hepatic mRNA levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2, as well as nitrite production and iNOS gene expression in both hepatocyte and macrophage cell lines. We demonstrated that EK100 exhibits potent protec-tion against hepatic IR injury, which may be used to design strategies to ameliorate liver damage during liver surgery.

Toxicity evaluation of water extract of tissue-cultured Taraxacum formosanum by acute, subacute administration, and Ames test

BACKGROUND: Taraxacum species (commonly known as dandelion) used as herbal medicine have been reported to exhibit an antiproliferative effect on hepatoma cells and antitumor activity in non-small-cell lung cancer cells. Although several investigations have demonstrated the safety of Taraxacum officinale, the safety of tissue-cultured plants of T. formosanum has not been assessed so far. Therefore, the present study examines the safety of the water extract of the entire plant of tissue cultured T. formosanum based on acute and subacute toxicity tests in rats, as well as the Ames tests. RESULTS: No death or toxicity symptoms were observed in the acute and subacute tests. The results of the acute test revealed that the LD50 (50% of lethal dose) value of the T. formosanum water extract for rats exceeded 5 g/kg bw. No abnormal changes in the body weight, weekly food consumption, organ weight, or hematological, biochemical, and morphological parameters were observed in the subacute toxicity test. Thus, the no observed adverse effect level (NOAEL) of T. formosanum water extract was estimated to be higher than 2.0 g/kg. Finally, the results of the Ames test revealed that T. formosanum water extract was not genotoxic at any tested concentration to any of five Salmonella strains. CONCLUSIONS: The water extract of tissue-cultured T. formosanum was non-toxic to rats in acute and subacute tests and exhibited no genotoxicity to five Salmonella strains.

Construction of a Lactobacillus plantarum Strain Expressing the Capsid Protein of Porcine Circovirus Type 2d (PCV2d) as an Oral Vaccine.

Porcine circovirus type 2 (PCV2) is a pathogenic virus that causes high rates of porcine death, resulting in severe economic losses to the swine industry. In recent years, the prevalence of PCV2d genotype infection in pigs has increased, but most commercially available vaccines were developed against the PCV2a strain and do not ensure complete protection from PCV2d. Here, we first constructed an expression vector for the antigenic ORF2-encoded capsid protein of PCV2d (pLp3050-His6-tag-capsid). We then utilized Lactobacillus plantarum to express the protein at mucosal sites in orally vaccinated mice. After transducing L. plantarum with pLp3050-His6-tag-capsid, the expressed protein could be found in cell wall and cell-free supernatant fractions by Western blotting. Using flow cytometry, we found that L. plantarum cells with surface-displayed capsid protein increased with time after SppIP induction. Finally, mice that were orally immunized 18 times with capsid-expressing L. plantarum showed increased levels of capsid-specific sIgA and virus neutralizing activity at mucosal sites, suggesting mucosal immunity had been stimulated by the vaccine. Overall, our findings demonstrate the feasibility and utility of a PCV2d-based vaccine, which may be of great value in porcine agriculture.

Adlay Bran Oil Suppresses Hepatic Gluconeogenesis and Attenuates Hyperlipidemia in Type 2 Diabetes Rats.

This study aimed to examine the antidiabetic effects of various concentrations of adlay bran oil (ABO) in high fat diet and streptozotocin-induced diabetic rats. Dietary supplementation with 10% ABO for 4 weeks effectively decreased the blood triacylglycerol, glucose, and total cholesterol levels in diabetic rats, although body weight remained the same. The mRNA and protein expressions of hepatic glucose transporter 2 (GLUT-2) and phosphoenolpyruvate carboxykinase (PEPCK) were increased and that of glucokinase (GCK) were decreased in diabetic rats. However, 10% ABO treatment reduced the mRNA and protein expressions of GLUT-2 and PEPCK and elevated the expression of hepatic GCK in diabetic rats. Thus, ABO enhanced hepatic glucose metabolism to decrease blood glucose in diabetic rats. In addition, 10% ABO supplementation increased the expression of phosphorylated protein kinase B (Akt) relative to the total Akt levels in the muscles of diabetic rats, indicating enhanced insulin sensitivity. The results indicate that ABO displays a potential for improving hyperlipidemia and hyperglycemia in diabetes by enhancing insulin sensitivity and hepatic glucose metabolism.

Use of electronic portal images to evaluate setup error and intra-fraction motion during free-breathing breast IMRT treatment.

Before delivering of intensity-modulated radiotherapy, kilo-voltage image-guidance radiotherapy is widely used in setup error correction and monitoring intra-fraction motion effectively. Accordingly, this study proposes and tests an image integration technique for observing intra-fraction motion during beam delivery, with the wider objective of reducing both image-guidance time and the dose delivered to normal breast tissue. The study sample comprised 33 female patients with breast cancer, and 241 sets of portal images acquired using a VARIAN aSi-1000 electronic portal imaging device. Motion amplitudes and vectors were collected and calculated separately by two senior therapists. The setup error in 3 axes was computed for every fraction, with average shifting for lateral, longitudinal and vertical direction was -0.3-mm ± 0.5, -0.1-mm ± 0.5 and -0.6-mm ± 1.6, with the average vector of setup error being 2.9-mm ± 1.4. The average intra-fraction motion for vertical direction was (A: -0.1-mm ± 1.0; B: -0.0 ± 1.1), for longitudinal was (A: -0.4-mm ± 1.7; B: 2.0 ± 1.1), and for lateral direction was (A: 0.3-mm ± 1.3; B: 0.2 ± 1.8). The average intra-fraction vector was 2.9-mm ± 1.3 for therapist A, and 3.4-mm ± 1.8 for therapist B. Offline Review commercial software was utilized for setup error and motion analysis, and data analysis and reliability testing were conducted with statistical package of the social sciences. Pearson correlations between the two therapists was moderate (0.59, p << 0.01), and the Cohen's kappa value for inter rater agreement between different evaluators was fair in the anterior-posterior direction (0.25, p << 0.01), with slight agreement in other two directions and vectors. The study presented efficient and dose reduction method to evaluate setup error and intra-fraction motion during breast intensity-modulated radiotherapy treatment.

Regulation of tumor progression via the Snail-RKIP signaling pathway by nicotine exposure in head and neck squamous cell carcinoma.

BACKGROUND: Recent studies suggest that long-term exposure of the carcinogen 4-methylnitrosamino-1-3-pyridyl-1-butanone (NNK) found in tobacco smoke is involved in the progression of head and neck squamous cell carcinoma (HNSCC). The underlying nicotine-mediated mechanism remains unclear. METHODS: An analysis of SCC-25 and Fadu cells with or without NNK exposure focusing on the evaluation of migration and invasion abilities, the expression of epithelial-mesenchymal transition, drug-resistance-related genes, properties of cancer stem cells (CSCs), and anti-apoptosis was performed. RESULTS: Long-term NNK exposure enhances migration and invasion with morphological alterations in a dose-dependently manner. Furthermore, NNK exposure also upregulates Snail, promotes sphere-forming ability, and overexpresses aldehyde dehydrogenase 1 (ALDH1), Nanog, OCT4, ABCG2, and MDR1. CONCLUSION: The current study confirmed that long-term NNK exposure plays a role in HNSCC by increasing anti-apoptosis and therapeutic resistance via the Snail-RKIP signaling pathway. Our data also suggest that α7 nicotinic acetylcholine receptor (α7-nAChR) inhibition or targeting Snail may provide a feasible rationale for preventing the progression of HNSCC.

Reciprocal relationship of Tn/NF-κB and sTn as an indicator of the prognosis of oral squamous cell carcinoma.

AIMS: In order to determine whether the expression of tumour-associated carbohydrate antigens (Tn/sTn) and a representative inflammation marker, nuclear factor-κB (NF-κB), is associated with the invasiveness of oral squamous cell carcinoma (OSCC), this study has attempted to investigate the correlation of the aforementioned markers with the well-established invasive pattern grading score (IPGS) and clinicopathological parameters. METHODS AND RESULTS: Specimens from 143 OSCC patients with classified clinicopathological parameters and IPGS were stained immunohistochemically using anti-Tn, sTn and NF-κB antibodies. Our results showed that the expression of both Tn and NF-κB was correlated positively with staging (P = 0.036; P = 0.015), recurrence (P < 0.001; P < 0.001) and distant metastasis (P = 0.005; P = 0.009), as well as with IPGS, while the expression of sTn was correlated inversely. In addition, poor survival was associated with overexpression of Tn and NF-κB but not with expression of sTn. CONCLUSIONS: Our results indicate that a reciprocal relationship between Tn and sTn expression may serve as a reliable indicator for OSCC prognostic evaluation. In addition, expression of Tn rather than sTn may play an important role in deeply invasive OSCC via regulation of NF-κB signalling.