Selenium nanoparticles decorated with polysaccharides from Sargassum fusiforme protects against 6-OHDA-induced neurotoxicity in PC12 cells and rat model of Parkinson's disease.

Parkinson's disease (PD) is a neurodegenerative disorder and identifying disease-causing pathways and drugs that target them has remained challenging. Herein, selenium nanoparticles decorated with polysaccharides from Sargassum fusiforme (SFPS-SeNPs) were investigated on 6-OHDA-induced neurotoxicity in PC12 cells and rats. 6-OHDA can significantly increase neurotoxicity, oxidative stress and decrease the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) both in vitro and vivo. In vitro, treatment with SFPS-SeNPs can significantly decrease 6-OHDA cytotoxicity, reactive oxygen species (ROS) production or malondialdehyde (MDA) levels, and cell apoptosis, significantly increased the activity of SOD and GPx. In vivo, 6-OHDA exposure could also decrease the expression of Nrf2 and OH-1, while treatment with SFPS-SeNPs (1 mg Se/kg) increased. SFPS-SeNPs can protect neurons from 6-OHDA-induced neurotoxicity by regulating apoptosis and Nrf2/ARE pathway. The present study demonstrated that SFPS-SeNPs is a good candidate for developing a new drug against neurodegenerative diseases such as PD.

Rapamycin alleviates irradiation-induced parotid injury by enhancing the whole gland homeostasis.

OBJECTIVES: Salivary gland injury is one of the most common complications of radiotherapy in head-and-neck cancers. This study investigated the mechanism by which rapamycin prevents irradiation (IR)-induced injury in the parotid glands. MATERIALS AND METHODS: Miniature pigs either received (a) no treatment (NT), (b) IR in the right parotid gland for 5 consecutive days (IR), or intraperitoneal administration of rapamycin (Rap) 1 h prior to IR (IR + Rap). Tissues were collected at three distinct time points (24 h, 4 weeks, and 16 weeks) after IR. Histological analyses, western blot, and real-time reverse transcriptase-polymerase chain reaction were performed to explore the mechanisms of IR-induced injury in the parotid gland. RESULTS: Rapamycin treatment maintained parotid salivary flow 16 weeks post-IR, preserved the number of acinar cells, and reduced parotid tissue fibrosis, as well as reduced apoptosis levels, decreased cleaved caspase-3 expression, and increased the Bcl-2/Bax ratio in the parotid glands. Autophagy marker LC3B was upregulated by rapamycin after IR, while P62 expression was downregulated. Rapamycin reduced the expression of pro-inflammatory factors and the mesenchymal tissue fibrosis following IR. CONCLUSIONS: Rapamycin maintains gland homeostasis after IR by decreasing apoptosis, reducing the expression of pro-inflammatory factors, and enhancing autophagy.

Effect of hydroxyethyl starch on drug stability and release of semaglutide in PLGA microspheres.

The degradation of peptide drugs limits the application of peptide drug microspheres. Structural changes of peptides at the water-oil interface and the destruction of their spatial structure in the complex microenvironment during polymer degradation can affect drug release and in vivo biological activity. This study demonstrates that adding hydroxyethyl starch (HES) to the internal aqueous phase (W1) significantly enhances the stability of semaglutide and optimizes its release behavior in PLGA microspheres. The results showed that this improvement was due to a spontaneous exothermic reaction (ΔH = -132.20 kJ mol-1) facilitated by hydrogen bonds. Incorporating HES into the internal aqueous phase using the water-in-oil-in-water (W1/O/W2) emulsion method yielded PLGA microspheres with a high encapsulation rate of 94.38 %. Moreover, microspheres with HES demonstrated well-controlled drug release over 44 days, unlike the slower and incomplete release in microspheres without HES. The optimized h-MG2 formulation achieved a more complete drug release (83.23 %) and prevented 30.65 % of drug loss compared to the HES-free microspheres within the same period. Additionally, the optimized semaglutide microspheres provided nearly three weeks of glycemic control with adequate safety. In conclusion, adding HES to the internal aqueous phase improved the in-situ drug stability and release behavior of semaglutide-loaded PLGA microspheres, effectively increasing the peptide drug payload in PLGA microspheres.

A highly sensitive Lock-Cas12a biosensor for detection and imaging of miRNA-21 in breast cancer cells.

The expression levels of microRNA (miRNA) vary significantly in correlation with the occurrence and progression of cancer, making them valuable biomarkers for cancer diagnosis. However, their quantitative detection faces challenges due to the high sequence homology, low abundance and small size. In this work, we established a strand displacement amplification (SDA) approach based on miRNA-triggered structural "Lock" nucleic acid ("Lock" DNA), coupled with the CRISPR/Cas12a system, for detecting miRNA-21 in breast cancer cells. The "Lock" DNA freed the CRISPR-derived RNA (crRNA) from the dependence on the target sequence and greatly facilitated the extended detection of different miRNAs. Moreover, the CRISPR/Cas12a system provided excellent amplification ability and specificity. The designed biosensor achieved high sensitivity detection of miRNA-21 with a limit of detection (LOD) of 28.8 aM. In particular, the biosensor could distinguish breast cancer cells from other cancer cells through intracellular imaging. With its straightforward sequence design and ease of use, the Lock-Cas12a biosensor offers significant advantages for cell imaging and early clinical diagnosis.

Analysis and prediction of 5-year survival in patients with cutaneous melanoma: a model-based period analysis.

Background: The survival and prognosis of patients are significantly threatened by cutaneous melanoma (CM), which is a highly aggressive disease. It is therefore crucial to determine the most recent survival rate of CM. This study used population-based cancer registry data to examine the 5-year relative survival rate of CM in the US. Methods: Period analysis was used to assess the relative survival rate and trends of patients with CM in the Surveillance, Epidemiology, and End Results (SEER) database during 2004-2018. And based on the data stratified by age, gender, race and subtype in the SEER database, a generalized linear model was 12established to predict the 5-year relative survival rate of CM patients from 2019 to 2023. Results: The 5-year relative survival increased to various degrees for both total CM and CM subtypes during the observation period. The improvement was greatest for amelanotic melanoma, increasing from 69.0% to 81.5%. The 5-year overall relative survival rates of CM were 92.9%, 93.5%, and 95.6% for 2004-2008, 2009-2013, and 2014-2018, respectively. Females had a marginally higher survival rate than males for almost all subtypes, older people had lower survival rates than younger people, white patients had higher survival rates than nonwhite ones, and urban locations had higher rates of survival from CM than rural locations did. The survival rate of CM was significantly lower for distant metastasis. Conclusion: The survival rate of patients with CM gradually improved overall during 2004-2018. With the predicted survival rate of 96.7% for 2019-2023, this trend will still be present. Assessing the changes experienced by patients with CM over the previous 15 years can help in predicting the future course of CM. It also provides a scientific foundation that associated departments can use to develop efficient tumor prevention and control strategies.

Screening of sensitive in vivo characteristics for early keratoconus diagnosis: a multicenter study.

Purpose: To analyze and compare sensitive in vivo characteristics for screening early keratoconus. Methods: This multicenter, case-control study included 712 eyes, after matching for age and biomechanically corrected intraocular pressure, from three clinics in different cities. The keratoconus (n = 288), early keratoconus (n = 91), and normal cornea (n = 333) groups included eyes diagnosed with bilateral keratoconus, fellow eyes with relatively normal topography with unilateral keratoconus, and normal eyes before refractive surgery, respectively. After adjusting for central corneal thickness, differences in vivo characteristics were analyzed among the three groups. The in vivo characteristics were measured by Pentacam and Corvis ST. Fifty-four indices were evaluated to screen for a sensitive index for the detection of early keratoconus. Results: Significant differences were observed in 26 of the 36 corneal biomechanical indeces between the early keratoconus and normal corneas. The area under the receiver operating characteristic curve of tomographic and biomechanical index, Belin/Ambrósio deviation, and Da in differentiating keratoconus from normal cornea was 1.000. Among the top five indeces of the area under the receiver operating characteristic curve for detecting early keratoconus, the corneal biomechanical-related index accounted for 80% (4/5), including A1 dArc length, highest concavity radius, A2 time, and tomographic and biomechanical index, of which the area under the receiver operating characteristic curve of A1 dArc length was 0.901. Conclusion: A1 dArc length and several corneal biomechanical indices are highly sensitive for the detection of early keratoconus, even in the absence of topographic abnormalities. Ophthalmologists should focus on the clinical application of corneal biomechanics and combine corneal tomography for the timely and accurate detection of early keratoconus.

Thermodynamic stability of cisplatin-loaded polymeric micelles and the phenotypic switching of the tumor-associated macrophages induced by combination of cisplatin-loaded micelles and Anti-PD-L1 antibody.

Chemotherapy is an effective anti-tumor treatment. Some anticancer chemotherapeutic drugs can not only induce cell death, but can also elicit antitumor immune responses. Here, the stability of cisplatin-loaded polymeric micelles (CDDP-PMs), pharmacokinetic drug-drug interactions of CDDP and anti-PD-L1 antibody (aPD-L1) in vivo and the alteration of the tumor microenvironment by combination of CDDP-PMs and aPD-L1 were evaluated. CDDP-PMs were fabricated by coordinated complexation and self-assembly method for tumor targeting. CDDP-PMs with higher mass ratio of copolymer have higher thermodynamic stability. The pharmacokinetic study showed that the CDDP and aPD-L1 were metabolized and cleared by two different pathways, suggesting that there is almost no risk of potential drug interactions between CDDP and aPD-L1 and the combination of aPD-L1 and CDDP- PMs may not alter the tissue distribution of CDDP. In vivo antitumor test showed that the tumor growth inhibition rates of CDDP-PMs combined with medium-dose aPD-L1 and CDDP-PMs combined with high-dose PD-L1 were 89.41% and 93.16%, respectively and therapeutic efficacy can be further increased by increasing the dose of aPD-L1 in co-administration group. This therapeutic system by combining chemotherapy and immunotherapy further increases the link between them and holds great potential to offer better safety and antitumor efficacy profiles.

Exercise downregulates HIPK2 and HIPK2 inhibition protects against myocardial infarction.

BACKGROUND: Exercise can protect myocardial infarction (MI) and downregulate cardiac Homeodomain-Interacting Protein Kinase 2 (HIPK2). However, the role of HIPK2 in MI is unclear. METHODS: HIPK2-/- mice and miR-222-/- rats, HIPK2 inhibitor (PKI1H) and adeno-associated virus serotype 9 (AAV9) carrying miR-222 were applied in the study. Animals were subjected to running, swimming, acute MI or post-MI remodeling. HIPK2 inhibition and P53 activator were used in neonatal rat cardiomyocytes (NRCMs) and human embryonic stem cell-derived cardiomyocytes (hESC-CMs) subjected to oxygen glucose deprivation/reperfusion (OGD/R). Serum miR-222 levels were analyzed in healthy people and MI patients that were survival or readmitted to the hospital and/or died. FINDINGS: Cardiac HIPK2 protein levels were reduced by exercise while increased in MI. In vitro, HIPK2 suppression by lentiviral vectors or inhibitor prevented apoptosis induced by OGD/R in NRCMs and hESC-CMs. HIPK2 inhibitor-treated mice and HIPK2-/- mice reduced infarct size after acute MI, and preserved cardiac function in MI remodeling. Mechanistically, protective effect against apoptosis by HIPK2 suppression was reversed by P53 activators. Furthermore, increasing levels of miR-222, targeting HIPK2, protected post-MI cardiac dysfunction, whereas cardiac dysfunction post-MI was aggravated in miR-222-/- rats. Moreover, serum miR-222 levels were significantly reduced in MI patients, as well as in MI patients that were readmitted to the hospital and/or died compared to those not. INTERPRETATION: Exercise-induced HIPK2 suppression attenuates cardiomyocytes apoptosis and protects MI by decreasing P-P53. Inhibition of HIPK2 represents a potential novel therapeutic intervention for MI. FUNDING: This work was supported by the grants from National Key Research and Development Project (2018YFE0113500 to JJ Xiao), National Natural Science Foundation of China (82020108002, 81722008, and 81911540486 to JJ Xiao, 81400647 to MJ Xu, 81800265 to YJ Liang), Innovation Program of Shanghai Municipal Education Commission (2017-01-07-00-09-E00042 to JJ Xiao), the grant from Science and Technology Commission of Shanghai Municipality (18410722200 and 17010500100 to JJ Xiao), the "Dawn" Program of Shanghai Education Commission (19SG34 to JJ Xiao), Shanghai Sailing Program (21YF1413200 to QL Zhou). JS is supported by Horizon2020 ERC-2016-COG EVICARE (725229).

Surface roughness effect on fatigue strength of aluminum alloy using revised stress field intensity approach.

The fatigue strength of a component is known to highly depend on its surface quality, and it is thus necessary to develop a reliable and appropriate mathematical model for fatigue strength assessment that consider the effect of surface roughness. In this paper, different underlying physical mechanisms of the roughness effect at different regions of specimens were studied by fatigue testing of 7N01 aluminum alloy. For a quantitative analysis of the surface roughness effect, a revised stress field intensity approach for a fatigue strength assessment of microsized notches was proposed as a theoretical support. In the new model, a new form of weight function was built to adapt the characteristics of microsized notches. In addition, the effect of the field radius was fundamentally weakened on solution of the stress field intensity and the difficulty of fatigue failure region definition in the traditional method was overcome correspondingly in the proposed model, which made the calculated field strength accurate and objective. Finally, to demonstrate the validity of the revised approach quantitatively, specimens with conventionally sized notches were subjected to stress field intensity calculations. The results showed that the revised approach has satisfactory accuracy compared with the other two traditional approaches from the perspective of quantitative analysis.

Overexpression of MsSiR enhances alkali tolerance in alfalfa (Medicago sativa L.) by increasing the glutathione content.

The sulfite reductase gene in Medicago sativa L. (MsSiR) encodes sulfite reductase (SiR) and catalyses the conversion of sulfite to sulfate in the sulfite assimilation pathway. In this study, we investigated the role of MsSiR in alfalfa by generating transgenic alfalfa that ectopically expressed MsSiR under the control of the CaMV35S promoter. The differences in alkali tolerance between the MsSiR-overexpressing and wild-type (WT) plants were analyzed, and the MsSiR-overexpressing plants exhibited an improved phenotype under alkali stress. Compared to WT plants, these plants demonstrated improved antioxidant activity as well as decreased H2O2 and O2- contents and increased glutathione reduced (GSH), Cysteine (Cys) and glutathione oxidized (GSSG) contents. MsSiR-overexpressing plants also exhibited high levels of adenosyl phosphosulfate reductases (APR), sulfite oxidase (SO) and MsSiR expression under alkali stress. It was speculated that MsSiR is involved in sulfur metabolism pathways, including the stabilization of sulfate and sulfite levels and the synthesis of GSH. These two processes achieve alkali tolerance by positively regulating the detoxification and antioxidant activities of alfalfa.

Sphingosine-1-phosphate alleviates irradiation-induced parotid injury in a miniature pig model.

OBJECTIVES: Our aim was to verify the alleviation effect of sphingosine-1-phosphate (S1P) in a miniature pig model. MATERIAL AND METHODS: Thirty male miniature pigs were randomly separated into 10 groups in our experiment. We administered S1P through the parotid duct in a retrograde fashion 2 hr before irradiation (IR). The salivary flow rate and blood flow rate were tested 20 weeks after IR. The apoptotic level was checked at 12, 24 hr and 7 days post-IR. RESULTS: Twenty weeks after IR, the salivary flow rate of the IR-side parotid gland in IR + S1P group can be maintained at about 40% of the non-IR side, while only 20% was maintained in the IR group. The blood flow rate and microvascular density were significantly higher in the IR + S1P group than in the IR group. The apoptotic level and cleaved caspase-3 expression were downregulated in IR + S1P group, and the ratio of Bcl-2/Bax was increased. The blood flow rate and CD31 level were significantly restored at 12, 24 hr and 7 days post-IR. CONCLUSION: Sphingosine-1-phosphate may partially alleviate IR-induced parotid dysfunction by decreasing apoptosis of microvascular endothelial cells and maintaining the blood flow rate.

A modified M-stage classification based on the metastatic patterns of pancreatic neuroendocrine neoplasms: a population-based study.

BACKGROUND: The present study aims to improve the M-stage classification of pancreatic neuroendocrine neoplasms (pNENs). METHODS: Two thousand six hundred sixty six pNENs were extracted from the Surveillance, Epidemiology, and End Results database to explore the metastatic patterns of pNENs. Metastatic patterns were categorized as single, two, or multiple (three or more) distant organ metastasis. The mean overall survival and hazard rate of different metastatic patterns were calculated by Kaplan-Meier and Cox proportional hazards models, respectively. The discriminatory capability of the modified M-stage classification was evaluated by Harrell's concordance index. RESULTS: The overall survival time significantly decreased with an increasing number of metastatic organs. In addition, pNENs with only liver metastasis had better prognosis when compared to other metastatic patterns. Thus, we modified the M-stage classification (mM-stage) as follows: mM0-stage, tumor without metastasis; mM1-stage, tumor only metastasized to liver; mM2-stage, tumor metastasized to other single distant organ (lung, bone, or brain) or two distant organs; mM3-stage, tumor metastasized to three or more distant organs. Harrell's concordance index showed that the modified M-stage classification had superior discriminatory capability than both the American Joint Committee on Cancer (AJCC) and the European Neuroendocrine Tumor Society (ENETS) M-stage classifications. CONCLUSIONS: The modified M-stage classification is superior to both AJCC and ENETS M-stage classifications in the prognosis of pNENs. In the future, individualized treatment and follow-up programs should be explored for patients with distinct metastatic patterns.

A surfactant-free microemulsion composed of isopentyl acetate, n-propanol, and water.

It has been demonstrated that in the absence of traditional surfactants, microemulsions can form from a ternary mixture of oil, water, and an amphi-solvent. These microemulsions are called surfactant-free microemulsions (SFMEs). To date, only a small number of SFME systems have been reported, and the current understanding of SFMEs is very limited. Herein, we report an SFME consisting of isopentyl acetate (IA), n-propanol, and water, in which IA (a simple ester compound) and n-propanol are used as the oil phase and amphi-solvent, respectively. The microstructures and structural transition of the SFME were investigated by cyclic voltammetry, fluorescence spectroscopy, and UV-visible spectroscopy techniques. Moreover, three kinds of microstructures, namely, oil-in-water (O/W), bicontinuous (BC), and water-in-oil (W/O), have been identified in the SFME, which are directly verified by cryo-TEM observations. A change in the composition of the SFME may lead to a structural transition from O/W through BC to W/O or vice versa, which is similar to the case of traditional surfactant-based microemulsions (SBMEs). To the best of our knowledge, this is the first time that the microstructures and structural transition of an SFME obtained using a simple ester compound as the oil phase have been identified.

Hypolipidaemic effect and mechanism of paprika seed oil on Sprague-Dawley rats.

BACKGROUND: Details regarding the functional properties of paprika seed oil are relatively scarce. In this study the hypolipidaemic effects and mechanisms of paprika seed oil on Sprague-Dawley rats are explored, which may improve the usage of paprika seed source and provide a theoretical basis of paprika seed oil for the alleviation of hyperlipidaemia. RESULTS: In capsaicin and paprika seed oil (PSO) groups, total cholesterol (TC) and total triglyceride (TG) in serum and liver lipids of rats were significantly decreased (P < 0.05). The contents of serum HDL cholesterol were increased and the contents of serum LDL cholesterol were decreased (P < 0.05). Real-time PCR analyses revealed that the hepatic mRNA expression of fatty acid synthetase (FAS) is decreased and the expression levels of HSL is increased (P < 0.05). The mRNA expression of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) is decreased and the expression levels of low-density lipoprotein receptor (LDLR) is significantly improved (P < 0.05). The cholesterol 7-hydroxylase (CYP7A1) expression is regulated to control the cholesterol-to-bile acid transformation and cholesterol excretion is promoted. Capsaicin and unsaturated fatty acid PSO can activate and improve the mRNA expression of transient receptor potential vanilloid type-1 (TRPV1) and peroxisome proliferators-activated receptors (PPARα). CONCLUSION: The hypolipidaemic effects of paprika seed oil (PSO) may be attributed to the inhibition of lipid synthesis via suppressing the expression of HMG-CoAR, CYP7A1 and FAS, meanwhile, promoting the metabolism and excretion of lipids via up-regulating the expression of LDLR, HSL, TRPV1 and PPARα. © 2017 Society of Chemical Industry.