Identification and characterization of murine adipose tissue-derived somatic stem cells of Shenque (CV8) acupoint.

BACKGROUND: Shenque (CV8) acupoint is located on the navel and has been therapeutically used for more than 2000 years in Traditional Chinese Medicine (TCM). However, clinical research on the underlying therapeutic molecular mechanisms of the CV8 acupoint lags far behind. This study aimed to study the mechanisms of umbilical acupoint therapy by using stem cells. METHODS: The morphological characteristics of CV8 acupoint were detected under a stereomicroscope using hematoxylin and eosin (H&E) staining. Oil Red, Masson, and immunohistochemical staining on multi-layered slices were used to identify the type of cells at the CV8 acupoint. Cell proliferation was measured by a cell counting kit-8 (CCK-8) method. Flow cytometry and immunohistochemistry were used for cell identification. Induced differentiation was used to compare the differentiation of cells derived from CV8 acupoint and non-acupoint somatic stem cells into other cell types, such as osteogenic, adipogenic, and neural stem cell-like cells. RESULTS: Morphological observations showed that adipose tissues at the linea alba of the CV8 acupoint in mice had a mass-like distribution. Immunohistochemical staining confirmed the distribution of stem cell antigen-1 (Sca-1) positive cells in the multi-layered slices of CV8 acupoint tissues. Cells isolated from adipose tissues at the CV8 acupoint exhibited high expression of Sca-1 and CD44 and low expression of CD31 and CD34, and these cells possessed osteogenic, adipogenic, and neurogenic stem cell-like cell differentiation ability. The cell proliferation (day 4: 0.5138 ±â€Š0.0111 vs. 0.4107 ±â€Š0.0180, t = 8.447, P = 0.0011; day 5: 0.6890 ±â€Š0.0070 vs. 0.5520 ±â€Š0.0118, t = 17.310, P < 0.0001; day 6: 0.7320 ±â€Š0.0090 vs. 0.6157 ±â€Š0.0123, t = 13.190, P = 0.0002; and day 7: 0.7550 ±â€Š0.0050 vs. 0.6313 ±â€Š0.0051, t = 42.560, P < 0.0001), adipogenic ([9.224 ±â€Š0.345]% vs. [3.933 ±â€Š1.800]%, t = 5.000, P = 0.0075), and neurogenic stem cell-like cell differentiation (diameter < 50 µm: 7.2000 ±â€Š1.3040 vs. 2.6000 ±â€Š0.5477, t = 7.273, P < 0.0001; diameter 50-100 µm: 2.6000 ±â€Š0.5477 vs. 1.0000 ±â€Š0.7071, t = 4.000, P = 0.0039; and diameter >100 µm: 2.6000 ±â€Š0.5477 vs. 0.8000 ±â€Š0.8367, t = 4.025, P = 0.0038) were significantly enhanced in somatic stem cells derived from the CV8 acupoint compared to somatic stem cells from the groin non-acupoint. However, cells possessed significantly weaker osteogenicity ([2.697 ±â€Š0.627]% vs. [7.254 ±â€Š0.958]%, t = 6.893, P = 0.0023) in the CV8 acupoint group. CONCLUSIONS: Our study showed that CV8 acupoint was rich with adipose tissues that contained abundant somatic stem cells. The biological examination of somatic stem cells derived from the CV8 acupoint provided novel insights for future research on the mechanisms of umbilical therapy.

Methods and Influencing Factors for the Simple and Rapid Identification of Depleted Uranium Weapon Use under Battlefield Conditions.

The purpose of this paper is to explore how to rapidly and easily identify depleted uranium (DU) samples under battlefield conditions and to study the factors that influence their measurement. The air-absorbed dose rate and surface contamination levels for DU samples of 2-330 g were measured using a patrol instrument and portable energy spectrometer. The results were analyzed in accordance with IAEA standards for judging radioactive substances. The energy spectra of 5-g quantities of DU samples were analyzed using a high-purity germanium gamma spectrometer, and the uranium content of 100 mg DU samples was determined with an inductively coupled plasma mass spectrometer to clarify the type and composition of the uranium. The same batches of DU samples were identified using a portable gamma-ray spectrometer. We added 0-5 g environmental soil powders at different proportions. After sealing, the spectra were collected with a detection distance of 1-5 cm for 10 min. The activities of U and U nuclides in the samples were detected with an NaI(TI) scintillation detector. The U and U mass abundances in samples were calculated from measured specific activities. The sample was determined to contain DU if the U to U ratio was below 0.00723. It is found that for detecting DU materials with a low activity, surface contamination level measurements are more effective than calculating the air-absorbed external irradiation dose rate. Hence, for low-activity samples suspected to be radioactive, a radiometer with a high sensitivity for surface contamination is recommended, and the optimal measurement distance is 1-3 cm. Under all detection conditions, U can be identified using a portable gamma spectrometer, whereas U can only be detected under certain conditions. If these nuclides can be detected simultaneously, a U to U ratio of below 0.00723 indicates the presence of DU. The main factors affecting this identification include the sample mass, sample purity, measurement distance, and measurement time. For the rapid identification of DU with a portable gamma-ray spectrometer, the mass of uranium in the sample must be more than 1 g, the measuring distance needs to be less than 1 cm, and the measuring time must be 1-10 min. It is feasible to use a portable gamma-ray spectrometer to rapidly identify the types and composition of nuclides in DU samples. The detection of U activity is a precondition for the identification of DU.

Nanoceria-Mediated Drug Delivery for Targeted Photodynamic Therapy on Drug-Resistant Breast Cancer.

Photodynamic therapy (PDT) has shown great potential for overcoming drug-resistant cancers. Here, we report a multifunctional drug delivery system based on chlorin e6 (Ce6)/folic acid (FA)-loaded branched polyethylenimine-PEGylation ceria nanoparticles (PPCNPs-Ce6/FA), which was developed for targeted PDT to overcome drug-resistant breast cancers. Nanocarrier delivery and FA targeting significantly promoted the cellular uptake of photosensitizers (PSs), followed by their accumulation in lysosomes. PPCNPs-Ce6/FA generated reactive oxygen species (ROS) after near-infrared irradiation (NIR, 660 nm), leading to reduced P-glycoprotein (P-gp) expression, lysosomal membrane permeabilization (LMP), and excellent phototoxicity toward resistant MCF-7/ADR cells, even at ultralow doses. Moreover, we identified NIR-triggered lysosomal-PDT using the higher dose of PPCNPs-Ce6/FA, which stimulated cell death by plasma membrane blebbing, cell swelling, and energy depletion, indicating an oncosis-like cell death pathway, despite the occurrence of apoptotic or autophagic mechanisms at lower drug doses. In vivo studies showed prolonged blood circulation times, low toxicity in mice, and high tumor accumulation of PPCNPs-Ce6/FA. In addition, using NIR-triggered PDT, PPCNPs-Ce6/FA displayed excellent potency for tumor regression in the MCF-7/ADR xenograft murine model. This study suggested that multifunctional PPCNPs-Ce6/FA nanocomposites are a versatile and effective drug delivery system that may potentially be exploited for phototherapy to overcome drug-resistant cancers, and the mechanisms of cell death induced by PDT should be considered in the design of clinical protocols.

Photosensitizer-Loaded Branched Polyethylenimine-PEGylated Ceria Nanoparticles for Imaging-Guided Synchronous Photochemotherapy.

A multifunctional theranostic platform based on photosensitizer (chlorin e6, Ce6)-loaded branched polyethylenimine-PEGylated ceria nanoparticles (PPCNPs-Ce6) was created for the development of effective cancer treatments involving the use of imaging-guided synchronous photochemotherapy. PPCNPs-Ce6 with high Ce6 photosensitizer loading (Ce6: cerium ∼40 wt %) significantly enhanced the delivery of Ce6 into cells and its accumulation in lysosomes, remarkably improving photodynamic therapeutic (PDT) efficacy levels compared to those in the administration of free Ce6 at ultralow drug doses (∼200 nM). Interestingly, PPCNPs-Ce6 efficiently induced HeLa cell death even at low concentrations (∼10 µM) without the use of laser irradiation and exhibit chemocytotoxicity. Inductively coupled plasma mass spectrometry (ICP-MS) and biology transmission electron microscopy (Bio-TEM) analyses demonstrated that ceria nanoparticles enter cells abundantly and accumulate in lysosomes or large vesicles. We then evaluated the effects of the different materials on lysosomal integrity and function, which revealed that PPCNPs-Ce6 catastrophically impaired lysosomal function compared to results with PPCNPs and Ce6. Studies of apoptosis revealed greater induction of apoptosis by PPCNPs-Ce6 treatment. This multifunctional nanocarrier also exhibited a high degree of solubility and stability in aqueous solutions, suggesting its applicability for extensive biomedical application.

PEGylated ceria nanoparticles used for radioprotection on human liver cells under γ-ray irradiation.

Ceria nanoparticles (CNPs) have recently been shown to protect cells and animals from radiation-induced damage. However, most of the CNPs used in previous studies were either naked or weakly protected by surfactants, which inevitably encounter many obstacles in biological applications. Here, alendronate was used as an ideal anchor to graft polyethylene glycol (PEG) onto CNPs, leading to enhanced stability, reduced cytotoxicity, and improved biological properties. Further investigation assessed the protective ability of the nanoparticles against radiation-induced effects for human normal liver cells (L-02), indicating that the PEGylated CNPs (CNPs-AL-PEG) were more efficient than naked CNPs. We determined that enhanced Ce(3+)/Ce(4+) ratios improved intracellular dispersion and that the ameliorated intracellular distribution of CNPs-AL-PEG contributes to the elevated expression of SOD2, which leads to increased protection of normal cells against ROS and reduces the oxidatively generated DNA damage. These studies hold tremendous promise for radioprotection and biological applications.

Interaction between maternal 5,10-methylenetetrahydrofolate reductase C677T and methionine synthase A2756G gene variants to increase the risk of fetal neural tube defects in a Shanxi Han population.

BACKGROUND: The 5,10-methylenetetrahydrofolate reductase (MTHFR) and methionine synthase (MS) are attractive candidates for screening for risk of neural tube defects (NTDs). The aim of the current study was to investigate maternal MTHFR and MS polymorphisms and the interaction between them and their influence on children with NTDs in the Shanxi Province of northern China. METHODS: Fifty-one mothers who previously had children with NTDs constituted the case group and 51 age-matched mothers with children that were unaffected by any birth defects constituted the control group. All subjects were genotyped for MTHFR C677T and MS A2756G polymorphisms. SPSS 11.5 software package was used for all analyses. RESULTS: There was a significant difference for MTHFR genotype distribution for one site (C677T) between the case and control groups. The T allele frequencies were significantly higher in the case group than in the control group (55.9% vs. 35.3%, P < 0.05). A lack of association was observed for the MS A2756G polymorphism. There was an interaction between the maternal MTHFR C677T genotype and MS A2756G genotype. CONCLUSION: Genetic interaction between MTHFR and MS genes raises the probability of neural tube defects.