Kidney-Targeted Drug Delivery System Based on Metformin-Grafted Chitosan for Renal Fibrosis Therapy.

Our previous study demonstrated that metformin plays an anti-fibrotic role in addition to its hypoglycemic effect. Worryingly, it often requires more than 5 times the hypoglycemic dose to achieve a satisfactory anti-fibrotic effect, which greatly increases the risk of systemic acidosis caused by metformin overdose. Low-molecular-weight chitosan (LMWC) has natural kidney-targeting properties and good biocompatibility and degradability. Thus, we synthesized a novel carrier metformin-grafted chitosan (CS-MET) based on an imine reaction between oxidized chitosan and metformin. Then, GFP was recruited to form GFP-loaded CS-MET nanoparticles (CS-MET/GFP NPs) with controllable particle size. We hypothesized that CS-MET/GFP NPs would enrich in the kidney and be absorbed by HK-2 cells via megalin-mediated endocytosis by intravenous injection, which may avoid systemic acidosis caused by metformin overdose. Subsequently, the nanoparticle ruptures and releases metformin to exert its anti-apoptotic, anti-inflammatory, and anti-fibrotic effects. Our results showed that CS-MET/GFP NPs have great transfection efficiency and could enter HK-2 cells mainly through megalin-mediated endocytosis. Compared to the free metformin, CS-MET/GFP NPs showed similar anti-apoptotic ability but better therapeutic effects on cellular inflammation and fibrosis in vitro. On the other hand, CS-MET/GFP NPs showed great kidney-targeting ability and superior anti-apoptotic, anti-inflammatory, and anti-fibrotic effects in vivo.

Post-discharge experiences of patients with extracorporeal membrane oxygenation support: A qualitative study.

PURPOSE: To explore the life experiences of patients who have been discharged after undergoing extracorporeal membrane oxygenation (ECMO) support. DESIGN: A qualitative descriptive approach was used. METHODS: Patients who have undergone ECMO support and have been discharged were recruited. Thirteen participants were involved in this study. The data were collected through a semi-structured interview and analyzed using the Colaizzi method. FINDINGS: Four major themes in life experiences were reported by the participants: changes in physical function, changes in psychological state, active adaptation to daily life, and substantial rehabilitation needs. CONCLUSION: Different, continuous, and convenient post-discharge physical and mental interventions, social support, spiritual support, and rehabilitation services should be provided according to the patient's circumstances. We also call on the government to increase the patient reimbursement rate for ECMO treatment. These measures may help to improve the quality of life of patients.

Codelivery of Doxorubicin and shAkt1 by Poly(ethylenimine)-Glycyrrhetinic Acid Nanoparticles To Induce Autophagy-Mediated Liver Cancer Combination Therapy.

Combination therapy has been developed as a promising therapeutic approach for hepatocellular carcinoma therapy. Here we report a low toxicity and high performance nanoparticle system that was self-assembled from a poly(ethylenimine)-glycyrrhetinic acid (PEI-GA) amphiphilic copolymer as a versatile gene/drug dual delivery nanoplatform. PEI-GA was synthesized by chemical conjugation of hydrophobic GA moieties to the hydrophilic PEI backbone via an acylation reaction. The PEI-GA nanocarrier could encapsulate doxorubicin (DOX) efficiently with loading level about 12% and further condense DNA to form PEI-GA/DOX/DNA complexes to codeliver drug and gene. The diameter of the complexes is 102 ± 19 nm with zeta potential of 19.6 ± 0.2 mV. Furthermore, the complexes possess liver cancer targeting ability and could promote liver cancer HepG2 cell internalization. Apoptosis of cells could be induced by chemotherapy of DOX, and PI3K/Akt/mTOR signaling pathway acts a beneficial effect on the modulation of autophagy. Here, it is revealed that utilizing PEI-GA/DOX/shAkt1 complexes results in effective autophagy and apoptosis, which are useful to cause cell death. The induction of superfluous autophagy is reported to induce type-II cell death and also could increase the sensity of chemotherapy to tumor cells. In this case, combining autophagy and apoptosis is meaningful for oncotherapy. In this study, PEI-GA/DOX/shAkt1 has demonstrated favorable tumor target ability, little side effects, and ideal antitumor efficacy.

Formulation and characterization of hydrophilic drug diclofenac sodium-loaded solid lipid nanoparticles based on phospholipid complexes technology.

To successfully prepare the diclofenac sodium (DS)-loaded solid lipid nanoparticles (SLNs), phospholipid complexes (PCs) technology was applied here to improve the liposolubility of DS. Solid lipid nanoparticles (SLNs) loaded with phospholipid complexes (PCs) were prepared by the modified emulsion/solvent evaporation method. DS could be solubilized effectively in the organic solvents with the existence of phospholipid and apparent partition coefficient of DS in PCs increased significantly. X-ray diffraction analysis suggested that DS in PCs was either molecularly dispersed or in an amorphous form. However, no significant difference was observed between the Fourier transform infrared spectroscopy (FT-IR) spectra of physical mixture and that of PCs. Particles with small sizes, narrow polydispersity indexes and high entrapment efficiencies could be obtained with the addition of PCs. Furthermore, according to the transmission electron microscopy, a core-shell structure was likely to be formed. The presence of PCs caused the change of zeta potential and retarded the drug release of SLNs, which indicated that phospholipid formed multilayers around the solid lipid core of SLNs. Both FT-IR and differential scanning calorimetry analysis also illustrated that some weak interactions between DS and lipid materials might take place during the preparation of SLNs. In conclusion, the model hydrophilic drug-DS can be formulated into the SLNs with the help of PCs.

Optimization of methazolamide-loaded solid lipid nanoparticles for ophthalmic delivery using Box-Behnken design.

The purpose of the present study was to optimize methazolamide (MTZ)-loaded solid lipid nanoparticles (SLNs) which were used as topical eye drops by evaluating the relationship between design factors and experimental data. A three factor, three-level Box-Behnken design (BBD) was used for the optimization procedure, choosing the amount of GMS, the amount of phospholipid, the concentration of surfactant as the independent variables. The chosen dependent variables were entrapment efficiency, dosage loading, and particle size. The generated polynomial equations and response surface plots were used to relate the dependent and independent variables. The optimal nanoparticles were formulated with 100 mg GMS, 150 mg phospholipid, and 1% Tween80 and PEG 400 (1:1, w/v). A new formulation was prepared according to these levels. The observed responses were close to the predicted values of the optimized formulation. The particle size was 197.8 ± 4.9 nm. The polydispersity index of particle size was 0.239 ± 0.01 and the zeta potential was 32.7 ± 2.6 mV. The entrapment efficiency and dosage loading were about 68.39% and 2.49%, respectively. Fourier transform infrared spectroscopy (FT-IR) study indicated that the drug was entrapped in nanoparticles. The optimized formulation showed a sustained release followed the Peppas model. MTZ-SLNs showed significant prolonged decreasing intraocular pressure effect comparing with MTZ solution in vivo pharmacodynamics studies. The results of acute eye irritation study indicated that MTZ-SLNs and AZOPT both had no eye irritation. Furthermore, the MTZ-SLNs were suitable to be stored at low temperature (4 °C).

In Silico Insights into Micro-Mechanism Understanding of Extracts of Taxus Chinensis Fruits Against Alzheimer's Disease.

BACKGROUND: The taxus chinensis fruit (TCF) shows promises in treatment of aging-related diseases such as Alzheimer's disease (AD). However, its related constituents and targets against AD have not been deciphered. OBJECTIVE: This study was to uncover constituents and targets of TCF extracts against AD. METHODS: An integrated approach including ultrasound extractions and constituent identification of TCF by UPLC-QE-MS/MS, target identification of constituents and AD by R data-mining from Pubchem, Drugbank and GEO databases, network construction, molecular docking and the ROC curve analysis was carried out. RESULTS: We identified 250 compounds in TCF extracts, and obtained 3,231 known constituent targets and 5,326 differential expression genes of AD, and 988 intersection genes. Through the network construction and KEGG pathway analysis, 19 chemicals, 31 targets, and 11 biological pathways were obtained as core compounds, targets and pathways of TCF extracts against AD. Among these constituents, luteolin, oleic acid, gallic acid, baicalein, naringenin, lovastatin and rutin had obvious anti-AD effect. Molecular docking results further confirmed above results. The ROC AUC values of about 87% of these core targets of TCF extracts was greater than 0.5 in the two GEO chips of AD, especially 10 targets with ROC AUC values greater than 0.7, such as BCL2, CASP7, NFKBIA, HMOX1, CDK2, LDLR, RELA, and CCL2, which mainly referred to neuron apoptosis, response to oxidative stress and inflammation, fibroblast proliferation, etc.Conclusions:The TCF extracts have diverse active compounds that can act on the diagnostic genes of AD, which deserve further in-depth study.

Automated spectrophotometric analyzer for rapid single-point titration of seawater total alkalinity.

An automated analyzer was developed to achieve fast, precise, and accurate measurements of seawater total alkalinity (AT) based on single-point titration and spectrophotometric pH detection. The single-point titration was carried out in a circulating loop, which allowed the titrant (hydrochloric acid and bromocresol green solution) and a seawater sample to mix at a constant volume ratio. The dissolved CO2 in the sample-titrant mixture was efficiently removed by an inline CO2 remover, which consists of a gas-permeable tubing (Teflon AF2400) submerged in a sodium hydroxide (NaOH) solution. The pH of the mixture was then measured with a custom-made spectrophotometric detection system. The analyzer was calibrated against multiple certified reference materials (CRMs) with different AT values. The analyzer features a sample throughput time of 6.5 min with high precision (±0.33-0.36 µmol kg(-1); n = 48) and accuracy (-0.33 ± 0.99 µmol kg(-1); n = 10). Intercomparison to a traditional open-cell AT titrator showed overall good agreement of 0.88 ± 2.03 µmol kg(-1) (n = 22). The analyzer achieved excellent stability without recalibration over 11 days, during which time 320 measurements were made with a total running time of over 40 h. Because of its small size, low power consumption requirements, and its ability to be automated, the new analyzer can be adapted for underway and in situ measurements.

Association of TLR4 gene 2026A/G (rs1927914), 896A/G (rs4986790), and 1196C/T (rs4986791) polymorphisms and cancer susceptibility: Meta-analysis and trial sequential analysis.

BACKGROUND: This study was performed to assess the association of TLR4 gene 2026A/G (rs1927914), 896A/G (rs4986790), and 1196C/T (rs4986791) polymorphisms and cancer susceptibility based on published case-control studies. METHODS: Web of Science, PubMed, Embase, CBM, WanFang Data, CNKI, and VIP database were used for article retrieving. Then, these articles were screened according to the study inclusion and exclusion criteria. The data was extracted, and the study quality was evaluated according to the principle of Newcastle-Ottawa Scale. Meta-analysis was performed by RevMan 5.4 and Stata MP-17 software. Trial sequential analysis was performed by TSA 0.9.5.10 Beta software. RESULTS: Eighty-seven case-control studies including 25,969 cases and 32,119 controls were included in the meta-analysis. The diseases involved in case groups include prostate cancer, lung cancer, gastric cancer, hepatocellular carcinoma, colorectal cancer, etc. A versus G model of rs1927914, A versus G model of rs4986790 and C versus T model of rs4986791 showed that odds ratio (OR) = 1.08, OR = 0.85, and OR = 0.74 respectively. All the 3 comparisons were statistically significant. Sensitivity analysis showed that the results were stable. Publication bias analysis and trial sequential analysis showed that no significant publication bias was found in the results of the meta-analysis, and the probability of false positives was small. CONCLUSION: People with A allele of rs1927914, G allele of rs4986790, or T allele of rs4986791 have higher risks of cancer. The results of meta-analysis are stable and have less probability of false positives.

Association of IL-27 gene rs153109 and rs17855750 polymorphisms with preeclampsia susceptibility and severity: Meta-analysis and trial sequential analysis.

BACKGROUND: The aim of this meta-analysis is to evaluate the association of interleukin-27 gene rs153109 and rs17855750 polymorphisms with preeclampsia susceptibility and severity. METHODS: Web of Science, PubMed, Embase, CBM, WanFang Data, CNKI, and VIP database were used for retrieving. After screening with our inclusion and exclusion criteria, data extraction and quantity evaluation were performed by 2 independent authors. Included case-control studies were used for meta-analysis by RevMan 5.4, and sensitivity analysis was carried out through 1-by-1 exclusion procedure. If heterogeneity exists, then random effects model was used; otherwise, fixed effect model was used. Publication bias analysis was performed using Begg test and Egger test. Trial sequential analysis was performed using trial sequential analysis 0.9.5.10 Beta. RESULTS: A total of 5 articles were included. The heterogeneity was high across most models during the meta-analysis. Meta-analysis results related to preeclampsia susceptibility showed that P values of all the models were higher than .05, while for meta-analysis results related to preeclampsia severity showed that P values of all the models were higher than .05 except for TT versus TG + GG and TT versus TG models of rs17855750 group. The sensitivity of the meta-analysis was high, and trial sequential analysis showed the possibility of false negative results. No obvious publication bias was found. CONCLUSIONS: There is no obvious association between interleukin-27 gene rs153109 and rs17855750 polymorphisms and preeclampsia susceptibility or severity. However, more multi-center and large sample case-control studies are expected to be carried out to verify our conclusion in the future.

Effect of nasal high-flow oxygen humidification on patients after cardiac surgery.

Background: Although high-flow humidified oxygen therapy (HFNC) has emerged as an important treatment for respiratory failure, few studies have reported on whether HFNC is appropriate for patients with hypoxemia after cardiac surgery, and the clinical efficacy of HFNC in patients undergoing cardiac surgery is unclear. Objective: To investigate the clinical effect of HFNC after cardiac surgery. Methods: Convenience sampling was used to select 76 patients who underwent invasive mechanical ventilation and oxygen therapy after valve replacement or coronary artery bypass grafting from July 2019 to June 2021. The patients were divided into the routine group and the HFNC group according to the oxygen therapy provided after the operation. The patients in the routine group (N = 38) were treated with oxygen inhalation by face mask after the operation, while those in the HFNC group (N = 38) were treated with HFNC via nasal cavity. The arterial partial pressure of oxygen (PaO2), the arterial partial pressure of carbon dioxide (PaCO2) and the oxygenation index (OI) were observed and compared between the two groups at 6 h, 12 h and 24 h after treatment. The sputum viscosity, incidence of second intubation and the intensive care unit (ICU) stay time were evaluated. Results: The difference in PaCO2 between the two groups was statistically significant at 24 h after treatment (p < 0.05). The PaO2 in the HFNC group was significantly higher than in the routine group at 24 h after treatment, and the OI of the routine group was lower than in the HFNC group at 6 h, 12 h and 24 h after treatment (p < 0.05). The sputum viscosity in the HFNC group was better than in the routine group at 12 h and 24 h after treatment. The second intubation rate and ICU stay time in the HFNC group were lower than in the routine group (p < 0.05). Conclusion: Compared with conventional mask oxygen inhalation, HFNC can effectively reduce sputum viscosity, improve oxygenation, reduce the incidence of repeated intubation and meet patients' comfort needs. It is an advantageous respiratory support strategy for patients after cardiac surgery compared with invasive mechanical ventilation to oxygen therapy and is beneficial to the recovery of cardiopulmonary function.

Uterine Ultrasound Doppler Hemodynamics of Magnesium Sulfate Combined with Labetalol in the Treatment of Pregnancy-Induced Hypertension Using Empirical Wavelet Transform Algorithm.

The aim of this study was to explore the hemodynamic changes of magnesium sulfate combined with labetalol in the treatment of pregnancy-induced hypertension (PIH) under Doppler uterine ultrasound based on the empirical wavelet transform (EWT) algorithm. 500 patients with PIH in the hospital were selected and randomly divided into the control group (n = 250) and the observation group (n = 250). The control group was treated with conventional magnesium sulfate; the observation group was given labetalol based on magnesium sulfate drip in the control group. The uterine artery blood flow simulation model was established based on the EWT algorithm and compared with a short-time Fourier transform (STFT). The normalized root mean square error (NRMSE) of the STFT method was 0.19, and the NRMSE extracted by the EWT method was 0.13. After treatment, the blood pressure index, 24-hour urinary protein, and incidence of adverse birth outcomes in the observation group were lower than those in the control group; the effective rate of the control group (90.4%) was lower than that of the observation group (97.6%); the hemodynamic indexes of the uterine artery in the observation group were lower than those in the control group, and the differences were statistically significant (P < 0.05). The estimation accuracy of the EWT method was higher than that of the traditional STFT method; the combined treatment of magnesium sulfate and labetalol in patients with PIH had a remarkable effect, which could control the blood pressure index and reduce the 24-hour urinary protein; the uterine artery Doppler ultrasound examination could change hemodynamics and improve the adverse outcomes of mothers and infants.

Development and application of "Special defibrillator for teaching and training".

BACKGROUND: To provide an economical and practical defibrillator for first aid teaching and training, to reduce the cost of teaching and training, increase teaching and training equipment, provide trainees with more hands-on training sessions, and improve first aid capabilities. METHODS: Developing a special teaching defibrillator with the same structure and operation configuration as the clinical medical emergency defibrillator. The appearance, structure and operating accessories of the two defibrillators are the same. The difference between the defibrillator and the clinical medical emergency defibrillator are as follows: the clinical medical emergency defibrillator can be energized, and there are expensive electronic accessories and defibrillation accessories for charging and discharging in the machine. When discharging, the electrode plate has current discharged into the human body; the power plug of the "special defibrillator for teaching and training" is a fake plug. When the power is plugged in, no current enters the body and the machine. There are no expensive electronic accessories and defibrillation accessories for charging and discharging, and no current is discharged during discharge. Then compare the teaching effect of the special defibrillator for teaching and training and the clinical medical emergency defibrillator (including operation score and attitude after training). RESULTS: The scores of defibrillator operation in the experimental group of junior college students (87.77 ± 4.11 vs. 83.30 ± 4.56, P < 0.001) and the experimental group of undergraduate students (90.40 ± 3.67 vs. 89.12 ± 3.68, P = 0.011) were higher than those in the corresponding control group; The attitude of junior college students in the experimental group and undergraduate students in the experimental group after training was more positive than that of the corresponding control group (P < 0.05). CONCLUSIONS: The special defibrillator for teaching and training can save the purchase cost of teaching equipment, increase teaching and training resources, and improve the trainee's defibrillation ability, defibrillation confidence and defibrillation security.

A potential new therapeutic system for glaucoma: solid lipid nanoparticles containing methazolamide.

Methazolamide (MTA) is an antiglaucoma drug; however, there are many side effects of its systemic administration with insufficient ocular therapeutic concentrations. The aim of this study was to formulate MTA-loaded solid lipid nanoparticles (SLNs) and evaluate the potential of SLNs as a new therapeutic system for glaucoma. SLNs were prepared by a modified emulsion-solvent evaporation method and their physicochemical characteristics were evaluated. The pharmacodynamics was investigated by determining the percentage decrease in intraocular pressure. The ocular irritation was studied by Draize test. Despite a burst release of SLNs, the pharmacodynamic experiment indicated that MTA-SLNs had higher therapeutic efficacy, later occurrence of maximum action, and more prolonged effect than drug solution and commercial product. Formulation of MTA-SLNs would be a potential delivery carrier for ocular delivery, with the advantages of a more intensive treatment for glaucoma, lower in doses and better patient compliance compared to the conventional eye drops.

Metformin attenuates renal tubulointerstitial fibrosis via upgrading autophagy in the early stage of diabetic nephropathy.

This study aimed at comparing the effects of metformin on tubulointerstitial fibrosis (TIF) in different stages of diabetic nephropathy (DN) in vivo and evaluating the mechanism in high glucose (HG)-treated renal tubular epithelial cells (RTECs) in vitro. Sprague-Dawley (SD) rats were used to establish a model of DN, and the changes of biochemical indicators and body weight were measured. The degree of renal fibrosis was quantified using histological analysis, immunohistochemistry, and immunoblot. The underlying relationship between autophagy and DN, and the cellular regulatory mechanism of metformin on epithelial-to-mesenchymal transition (EMT) were investigated. Metformin markedly improved renal function and histological restoration of renal tissues, especially in the early stages of DN, with a significant increase in autophagy and a decrease in the expression of fibrotic biomarkers (fibronectin and collagen I) in renal tissue. Under hyperglycemic conditions, renal tubular epithelial cells inactivated p-AMPK and activated partial EMT. Metformin-induced AMPK significantly ameliorated renal autophagic function, inhibited the partial EMT of RTECs, and attenuated TIF, all of which effectively prevented or delayed the onset of DN. This evidence provides theoretical and experimental basis for the following research on the potential clinical application of metformin in the treatment of diabetic TIF.

Association of Cytochrome P450 2C9*3 and Angiotensin II Receptor 1 (1166A>C) Gene Polymorphisms With the Antihypertensive Effect of Irbesartan.

BACKGROUND: To analyze whether the cytochrome P450 enzyme 2C9*3 (CYP2C9*3) and angiotensin II receptor 1 (AGTR1) (1166A>C) gene polymorphisms are associated with the risk of essential hypertension (EH) and the antihypertensive effect of irbesartan. METHODS: A total of 2,057 EH patients and 286 healthy controls were enrolled for genotyping in which 598 EH patients were given irbesartan 150 mg/day for 4 weeks. Blood pressure of all subjects were determined before and at the end of 4-week treatment. RESULTS: There was no significant difference in genotype frequencies of CYP2C9*3 and AGTR1 (1166A>C) between EH and control groups. Subjects with *1*3/*3*3 genotypes of the CYP2C9*3 gene had larger systolic and diastolic blood pressure reductions (34.9 ± 15.5 vs. 29.3 ± 10.2 mm Hg and 22.8 ± 9.0 vs. 19.6 ± 8.5 mm Hg, respectively) compared with the *1*1 genotype. For AGTR1 (1166A>C) polymorphisms, although there was no significant difference among AC, CC, and AA genotypes, male subjects with AC/CC genotypes had larger systolic and diastolic blood pressure reductions (32.3 ± 1.3 vs. 29.3 ± 0.5 mm Hg and 21.6 ± 0.8 vs. 19.4 ± 0.1 mm Hg, respectively, P < 0.05) in response to irbesartan treatment compared with the AA genotype. CONCLUSIONS: Polymorphisms of CYP2C9*3 and AGTR1 (1166A>C) are not significantly different between EH and healthy controls. Male subjects with AC and CC genotypes of AGTR1 (1166A>C) show better antihypertensive effect of irbesartan than the AA genotype.

Gene therapy strategies for rare monogenic disorders with nuclear or mitochondrial gene mutations.

Rare monogenic disorders are a group of single-gene-mutated diseases that have a low incidence rate (less than 0.5‰) and eventually lead to patient disability and even death. Due to the relatively low number of people affected, these diseases typically fail to attract a great deal of commercial investment and research interest, and the affected patients thus have unmet medical needs. Advances in genomics biology, gene editing, and gene delivery can now offer potentially effective options for treating rare monogenic diseases. Herein, we review the application of gene therapy strategies (traditional gene therapy and gene editing) against various rare monogenic diseases with nuclear or mitochondrial gene mutations, including eye, central nervous system, pulmonary, systemic, and blood cell diseases. We summarize their pathologic features, address the barriers to gene delivery for these diseases, discuss available therapies in the clinic and in clinical trials, and sum up in-development gene delivery systems for various rare monogenic disorders. Finally, we elaborate the possible directions and outlook of gene therapy for rare monogenic disorders.

Assessing the efficiency of constructed wetlands in removing PPCPs from treated wastewater and mitigating the ecotoxicological impacts.

The prevalence of pharmaceuticals and personal care products (PPCPs) in municipal wastewater has led to increased concerns about their impact on both human health and ecosystem. The constructed wetlands have been recognized as one of the cost-effective and green mitigation approaches to remove the PPCPs in the municipal wastewater. In this study, the effectiveness of a full scale constructed wetlands treatment system (CCWTs) in removing the 36 PPCPs was investigated. The load mass of PPCPs discharged by the wastewater treatment plant into the CCWTs was calculated. Removal efficiencies of PPCPs were evaluated based on physico-chemical properties such as octanol-water partition coefficient (Log kow), molecular weight (MW, g mol-1) and the acid dissociation constant (pKa). The CCWTs are especially efficient in removing azithromycin, sertraline, tolfenamic acid, and diphenhydramine with removing efficiency >88%. However, the removal efficiencies of PPCPs in CCWTs exhibit a large variability, depending on physical and chemical properties of the molecules, with 4.7-96.7% for antibiotics, 5-86% for antidepressant and antiseizure drugs, 3.5-88% for NSAIDs, 29-77% for ß-blockers and statins and 5.5-94% for other types of PPCPs. In addition, the environmental risk assessment showed that majority of the PPCPs (excluding sulfamethoxazole) in the effluent yielded low aquatic risk (risk quotient, RQ ≤ 0.1) due to the efficiency of CCWTs. The toxicity index scores were calculated by integration of the predicted and available toxicological hazard data into the prioritization ranking algorithm through Toxicological Prioritization Index (ToxPi).

Preparation and evaluation of in situ gelling ophthalmic drug delivery system for methazolamide.

PURPOSE: In this study, a thermosensitive in situ gelling vehicle was prepared to increase the precorneal resident time and the bioavailability of methazolamide (MTA). METHOD: Poloxamer analogs were used as the gelling agents, and the in situ gel was obtained by using a cold method. The gelation temperature, rheological properties, in vitro release as well as in vivo evaluation (the elimination of MTA in aqueous humor and intraocular-lowering effect) of the optimized formulations were investigated. RESULTS: The optimum concentrations of poloxamer analogs for the in situ gel-forming delivery system were 21% (w/w) poloxamer 407 and 10% (w/w) poloxamer P188. This formulation was able to flow freely under nonphysiological conditions and underwent sol-gel transition in the cul-de-sac upon placement into the eye. In vitro release studies demonstrated a diffusion-controlled release of MTA from the poloxamer solutions over a period of 10 hours. In vivo evaluation indicated that the poloxamer solutions had a better ability to retain drug than MTA eyedrops did. CONCLUSION: These results suggested that in situ gelling ophthalmic drug delivery system may hold some promise in ocular MTA delivery.

Exosomes derived from GDNF-modified human adipose mesenchymal stem cells ameliorate peritubular capillary loss in tubulointerstitial fibrosis by activating the SIRT1/eNOS signaling pathway.

Mesenchymal stem cells (MSCs) have emerged as ideal cell-based therapeutic candidates for the structural and functional restoration of the diseased kidney. Glial cell line-derived neurotrophic factor (GDNF) has been demonstrated to promote the therapeutic effect of MSCs on ameliorating renal injury. The mechanism may involve the transfer of endogenous molecules via paracrine factors to salvage injured cells, but these factors remain unknown. Methods: GDNF was transfected into human adipose mesenchymal stem cells via a lentiviral transfection system, and exosomes were isolated (GDNF-AMSC-exos). Using the unilateral ureteral obstruction (UUO) mouse model and human umbilical vein endothelial cells (HUVECs) against hypoxia/serum deprivation (H/SD) injury models, we investigated whether GDNF-AMSC-exos ameliorate peritubular capillary (PTC) loss in tubulointerstitial fibrosis and whether this effect is mediated by the Sirtuin 1 (SIRT1) signaling pathway. Additionally, by using SIRT1 activators or siRNAs, the roles of the candidate mRNA and its downstream gene in GDNF-AMSC-exo-induced regulation of endothelial cell function were assessed. PTC characteristics were detected by fluorescent microangiography (FMA) and analyzed by the MATLAB software. Results: The green fluorescent PKH67-labeled exosomes were visualized in the UUO kidneys and colocalized with CD81. GDNF-AMSC-exos significantly decreased PTC rarefaction and renal fibrosis scores in mice with UUO. In vitro studies revealed that GDNF-AMSC-exos exerted cytoprotective effects on HUVECs against H/SD injury by stimulating migration and angiogenesis as well as conferring apoptosis resistance. Mechanistically, GDNF-AMSC-exos enhanced SIRT1 signaling, which was accompanied by increased levels of phosphorylated endothelial nitric oxide synthase (p-eNOS). We also confirmed the SIRT1-eNOS interaction in HUVECs by immunoprecipitation. Furthermore, we observed a correlation of the PTC number with the SIRT1 expression level in the kidney in vivo. Conclusion: Our study unveiled a mechanism by which exosomes ameliorate renal fibrosis: GDNF-AMSC-exos may activate an angiogenesis program in surviving PTCs after injury by activating the SIRT1/eNOS signaling pathway.

miR-96 Inhibits SV2C to Promote Depression-Like Behavior and Memory Disorders in Mice.

Accumulating evidence continues to emphasize the role of microRNAs as significant contributors to depression-like behavior and memory disorders. The current study aimed to investigate the mechanism by which miR-96 influences depression-like behavior and memory deficit in mice. A depression-like behavior and memory disorder mouse model was initially established by means of intraperitoneal injection with lipopolysaccharide. Memory deficits in the mice were evaluated using the Novel Object Recognition Test and Morris water maze experiments, whereas the Sucrose Preference Experiment and forced swimming experiments were performed to identify depression-like behavior in mice. The levels of tumor necrosis factor-α, malondialdehyde, superoxide dismutase, glutathione, and the monoamine transmitters 5-hydroxytryptamine and dopamine were subsequently detected in the serum. Reverse transcription-quantitative polymerase chain reaction and Western blot analysis evaluated the expression of miR-96 and SV2C expression in the CA1 hippocampal region of the mice. Finally, the relationship of miR-96 and SV2C was verified by dual-luciferase reporter gene assay. Our data indicated that the expression of miR-96 was increased, whereas that of SV2C was decreased in the CA1 region of mice exhibiting depression-like behavior and memory impairment. When miR-96 was downregulated or SV2C was overexpressed via intra-cerebroventricular injection with a miR-96 antagonist (miR-96 antagomir) or overexpression of SV2C vector, the Novel Object Recognition Test and sucrose preference index were increased, whereas the escape latency, the number of water maze platform crossings, and the immobility time of the mice were decreased. The serum levels of tumor necrosis factor-α, interleukin-1ß, and malondialdehyde in the mouse CA1 region of mice were reduced, whereas the levels of superoxide dismutase and glutathione were elevated after the downregulation of miR-96 or overexpression of SV2C. Collectively, our study demonstrates that miR-96 negatively regulates the expression of SV2C, which consequently leads to depression-like behavior and memory impairment in mice. Our findings highlight the potential of miR-96-targeted therapeutics.