Function and biomedical implications of exosomal microRNAs delivered by parenchymal and nonparenchymal cells in hepatocellular carcinoma.

Small extracellular vesicles (exosomes) are important components of the tumor microenvironment. They are small membrane-bound vesicles derived from almost all cell types and play an important role in intercellular communication. Exosomes transmit biological molecules obtained from parent cells, such as proteins, lipids, and nucleic acids, and are involved in cancer development. MicroRNAs (miRNAs), the most abundant contents in exosomes, are selectively packaged into exosomes to carry out their biological functions. Recent studies have revealed that exosome-delivered miRNAs play crucial roles in the tumorigenesis, progression, and drug resistance of hepatocellular carcinoma (HCC). In addition, exosomes have great industrial prospects in the diagnosis, treatment, and prognosis of patients with HCC. This review summarized the composition and function of exosomal miRNAs of different cell origins in HCC and highlighted the association between exosomal miRNAs from stromal cells and immune cells in the tumor microenvironment and the progression of HCC. Finally, we described the potential applicability of exosomal miRNAs derived from mesenchymal stem cells in the treatment of HCC.

[Analysis on applications and funded projects in physiology and integrative biology of National Natural Science Foundation of China in the past decade].

Here we perform a review on applications and funded projects at Division of Physiology and Integrative Biology in Department of Life Sciences sponsored by National Natural Science Foundation of China in the past ten years. Based on the research fields of applications and funded projects and the funding cost, we analyzed the sub-disciplines of the funded applications, key support areas, research frontiers and trends in the subjects of physiology and integrative biology, which gives us an insight into the future applications to optimize the layout of research areas in Division of Physiology and Integrative Biology.

[IFTIR study on the micro-structure in the construction process of water-borne coatings].

Borne paint was studied in the article in which the solvent water was regarded as a variable factor. A series of paint samples with different percentage of water were configured before observing their storage performance and microstructures by using Fourier-transform infrared (FTIR) ATR (attenuated total reflection) spectroscopy and diffuse reflectance (DF) FTIR spectroscopy. The effects of construction process and solvent water on the coating film were examined through analyzing the changes of internal functional groups before and after coating. The results indicated that as the amount of solvent water was increased, the characteristic peak of nu(C=O) in 1 727 cm(-1) turned into acromion, the absorption peakin 871 cm(-1) was blue shifted, the intensity of the peak of hydroxyl (nu(OH)) in 3400 cm(-1) and unsaturated absorption peak of nu-CH in 3030 cm(-1) gradually weakened, the absorption peaks strengths of nu-CH3 and nu-CH2 in 2962 and 2871 cm(-1) increased, respectively. The absorption peak in 2516 cm(-1) was shifted to 2603 cm(-1) then form the acromion, the absorption peak of 1647 cm(-1) gradually changed to 1455 cm(-1), the stretching vibration absorption peak of nu(C-O) in 1107 cm(-1) was red-shifted. During the coating process of paint, the solvent water and paint molecules interacted with each other, having a significant impact on the electron cloud density distribution of the paint molecules. The results obtained in the article have an important significance for the production and construction of paint.

[Investigation of FTIR spectra analysis of gallstones from Xuzhou region by solvent extraction].

The present paper is a preliminary exploration of the possible way the gallstones are formed. Five categories of gallstones from clinical surgery in Xuzhou region were extracted by a series of solvents. Fourier transform infrared spectroscopy (FTIR) was used to characterize the structure of morphological changes between gallstone and residue by extracting. The gallstone samples are from clinical surgeries in Xuzhou region where gallstone disease is quite common. Samples were extracted with a series of solvents, and then FTIR and other instrumental analysis were applied to characterize the composition, structure and morphological changes of the samples both before and after the extraction. The results show that the gallstone samples can be classified as 5 types: cholesterol-type, cholesterol-based hybrid type with salt, bilirubin and protein as its insoluble substances, brown pigment type and black pigment type gallstones. The results also indicate that protein plays a key role in gallstone nucleation process by providing a sediment matrix for the formation of gallstones. Both cholesterol and carbonated hydroxyapatite are found in the insoluble substances of the samples. It is possible that cholesterol was accompanied by carbonated hydroxyapatite and there are interactions between them, and these interactions contribute to the crystallization process and speed up the formation of gallstones. All the results above may provide useful references for the clinical diagnosis, treatment and prevention of gallstones.

[Investigation of FTIR spectra analysis on carbon dioxide absorption with improved amine solution].

Carbon dioxide is a major sort of greenhouse gas as well as important carbon resource. With the developments of industries, emission of carbon dioxide has increased sharply. Hence, controls of carbon dioxide emission and resource transformation have become the hotspot of current study. As a new kind of carbon resource, the fields of CO2 research and application are very extensive. Among those methods, the amine absorption has good qualities of faster absorption rate, higher efficiency and so on, so it has been widely studied. But organic amine have such shortcomings: high consumption of heat energy, strong corrosive and easy oxidated, now pursuer mainly focused on the organic amine modified. The results showed that, when the time the amount of antioxidant 1010 is 0.152, the absorption capacity is 2503.53 mL. the volume of analysis is 982.00 mL, and the absorption rate changes more slowly, by FTIR, Samples of its renewable-OH associating is not apparent that the antioxidant content in 1010, oxidation products of the MEA is acid or less oxidation and antioxidant 1010 product in early to respond fully to form stable non-radical compounds. Therefore, the best dosage of antioxidant 1010 is 0.15%. When the time that the amount of Na2SO3 is 0.15%, the absorption capacity is 2922.88 mL. Analysis of the volume is 723.00 mL, by FTIR, which reveals the oxidation products of the MEA is amide -C=O which in alkaline solution can be transiting into primary amine, and be easy absorbing CO2. Comparing the antioxygenic proerty of antioxidant 1010 with Na2SO3, from the absorption rate, the amount of absorption , Na2SO3's antioxidant properties is superior than antioxidant 1010; by infrared spectral analysis, 1010/20% MEA solution's oxidation products is the acid, Na2SO3/20% MEA solutions, the oxidation product is amide, amide solution is advantaged for absorbing CO2, So Na2SO3's antioxidant properties is superior than antioxidant 1010.

Atrial natriuretic peptide regulates Ca channel in early developmental cardiomyocytes.

BACKGROUND: Cardiomyocytes derived from murine embryonic stem (ES) cells possess various membrane currents and signaling cascades link to that of embryonic hearts. The role of atrial natriuretic peptide (ANP) in regulation of membrane potentials and Ca(2+) currents has not been investigated in developmental cardiomyocytes. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the role of ANP in regulating L-type Ca(2+) channel current (I(CaL)) in different developmental stages of cardiomyocytes derived from ES cells. ANP decreased the frequency of action potentials (APs) in early developmental stage (EDS) cardiomyocytes, embryonic bodies (EB) as well as whole embryo hearts. ANP exerted an inhibitory effect on basal I(CaL) in about 70% EDS cardiomyocytes tested but only in about 30% late developmental stage (LDS) cells. However, after stimulation of I(CaL) by isoproterenol (ISO) in LDS cells, ANP inhibited the response in about 70% cells. The depression of I(CaL) induced by ANP was not affected by either Nomega, Nitro-L-Arginine methyl ester (L-NAME), a nitric oxide synthetase (NOS) inhibitor, or KT5823, a cGMP-dependent protein kinase (PKG) selective inhibitor, in either EDS and LDS cells; whereas depression of I(CaL) by ANP was entirely abolished by erythro-9-(2-Hydroxy-3-nonyl) adenine (EHNA), a selective inhibitor of type 2 phosphodiesterase(PDE2) in most cells tested. CONCLUSION/SIGNIFICANCES: Taken together, these results indicate that ANP induced depression of action potentials and I(CaL) is due to activation of particulate guanylyl cyclase (GC), cGMP production and cGMP-activation of PDE2 mediated depression of adenosine 3', 5'-cyclic monophophate (cAMP)-cAMP-dependent protein kinase (PKA) in early cardiomyogenesis.

[Periodic and fractal precipitation in ATP-Co(2+)-decoxycholate gel system].

In the simulation experiments in vitro of the formation of gallstone, adenosine-triphosphate(ATP)-Co(2+)-deoxycholic acid(DC) gel system was chosen to study the periodic precipitation progress. The effect of ATP on the Co(2+)-DC gel system was also determined, and the structure of the periodic precipitation formed was characterized by FTIR. The results show that the patterns formed in the systems with ATP are different, ATP affected the rate and structure of precipitation through its variable participation in the metal coordination complexes as judged by the phosphate P=O bands and the deoxycholate COO- symmetric and asymmetric vibration bands as measured by FTIR Theses spectroscopic differences were correlated with color and pattern differences in the precipitates. ATP has a more remarkable function than AMP to the modes of patterns, meanwhile the system patterns transform from fractal to periodic precipitation. There is a complex interaction among ATP, sodium deoxycholic and Co2+ with a transparent crystal produced. The crystal is deoxycholic acid and the periodic precipitation is composed of ATP and DC covalent to Co2+. These results indicate that stone formation and remodeling is a dynamic, nonlinear progress. Much of the precipitate, as judged by local differences in composition, is not in equilibrium with the general gel environment. The authors conclude that the formation of gallstone features complex and nonlinear chemical character, in which nucleotides as living material play a very important role.

Improvement of SSO-mediated gene repair efficiency by nonspecific oligonucleotides.

Targeted gene repair mediated by single-stranded DNA oligonucleotides (SSOs) is a promising method to correct the mutant gene precisely in prokaryotic and eukaryotic systems. We used a HeLa cell line, which was stably integrated with mutant enhanced green fluorescence protein gene (mEGFP) in the genome, to test the efficiency of SSO-mediated gene repair. We found that the mEGFP gene was successfully repaired by specific SSOs, but the efficiency was only approximately 0.1%. Then we synthesized a series of nonspecific oligonucleotides, which were single-stranded DNA with different lengths and no significant similarity with the SSOs. We found the efficiency of SSO-mediated gene repair was increased by 6-fold in nonspecific oligonucleotides-treated cells. And this improvement in repair frequency correlated with the doses of the nonspecific oligonucleotides, instead of the lengths. Our evidence suggested that this increased repair efficiency was achieved by the transient alterations of the cellular proteome. We also found the obvious strand bias that antisense SSOs were much more effective than sense SSOs in the repair experiments with nonspecific oligonucleotides. These results provide a fresh clue into the mechanism of SSO-mediated targeted gene repair in mammalian cells.

Nitric oxide mediates stretch-induced Ca2+ release via activation of phosphatidylinositol 3-kinase-Akt pathway in smooth muscle.

BACKGROUND: Hollow smooth muscle organs such as the bladder undergo significant changes in wall tension associated with filling and distension, with attendant changes in muscle tone. Our previous study indicated that stretch induces Ca(2+) release occurs in the form of Ca(2+) sparks and Ca(2+) waves in urinary bladder myocytes. While, the mechanism underlying stretch-induced Ca2+ release in smooth muscle is unknown. METHODOLOGY/PRINCIPAL FINDINGS: We examined the transduction mechanism linking cell stretch to Ca(2+) release. The probability and frequency of Ca(2+) sparks induced by stretch were closely related to the extent of cell extension and the time that the stretch was maintained. Experiments in tissues and single myocytes indicated that mechanical stretch significantly increases the production of nitric oxide (NO) and the amplitude and duration of muscle contraction. Stretch-induced Ca(2+) sparks and contractility increases were abrogated by the NO inhibitor L-NAME and were also absent in eNOS knockout mice. Furthermore, exposure of eNOS null mice to exogenously generated NO induced Ca(2+) sparks. The soluble guanylyl cyclase inhibitor ODQ did not inhibit SICR, but this process was effectively blocked by the PI3 kinase inhibitors LY494002 and wortmannin; the phosphorylation of Akt and eNOS were up-regulated by 204+/-28.6% and 258+/-36.8% by stretch, respectively. Moreover, stretch significantly increased the eNOS protein expression level. CONCLUSIONS/SIGNIFICANCE: Taking together, these results suggest that stretch-induced Ca2+ release is NO dependent, resulting from the activation of PI3K/Akt pathway in smooth muscle.

[FTIR investigation of organic phase containing rare earth ion in naphthenic acid-phosphonate ester-heptane system].

Naphthenic acid (NA) was mixed with PC88A in heptane, and 10 moL x L(-1) KOH aqueous solution was used to saponify the solution subsequently. The transparent and clear appearance of the obtained solution indicated the formation of w/o micell and microemusion. In the present study, micell with high saponification percentage (80%) was used to extract neodymium. The organic phase structure was characterized using FTIR spectroscopy in the extraction process. IR spectra indicate that the characteristic peaks of nu(COOH), nu(s)(COO-) and nu(P=O) shift to lower wavenumbers as the loading of Nd in organic phase increases. The results demonstrated that Nd(III) are coordinated to P=O and COO- groups simultaneously, and the supramolecules with self-assembly structure was formed containing PC88A and NA, which were binding with Nd(III) in the extracted organic phase.

[Study on the structure of periodic and chaotic patterns of AMP-DC-Cu-gel systems].

This is an in vitro model to mimic the conditions present during gallstone formation. Adenosine mono-phosphate (AMP), an important bio-molecule, was chosen. Its effect on the formation of periodic/chaotic patterns in the deoxycholate-CuCl2-gel and deoxycholate-CuCl2-glucose-gel systems were studied. FTIR spectroscopy was used to identify the structure of the precipitates. The results indicate that the patterns formed in the systems with AMP are different. And FTIR spectra show that AMP may also be involved in the coordination sphere of Cu2+ ion. The results are more complicated in the deoxycholate-CuCl2 system containing glucose, suggesting that the nonlinear scientific concept should be considered in understanding gallstone formation.

Targeted correction of a chromosomal point mutation by modified single-stranded oligonucleotides in a GFP recovery system.

Synthetic oligonucleotides had been employed in DNA repair and promised great potentials in gene therapy. To test the ability of single-stranded oligonucleotide (SSO)-mediated gene repair within a chromosomal site in human cells, a HeLa cell line stably integrated with mutant enhanced green fluorescence protein gene (mEGFP) in the genome was established. Transfection with specific SSOs successfully repaired the mEGFP gene and resulted in the expression of functional fluorescence proteins, which could be detected by fluorescence microscopy and FACS assay. Western blot showed that EGFP was only present in the cells transfected with correction SSOs rather than the control SSOs. Furthermore, DNA sequencing confirmed that phenotype change resulted from the designated nucleotide correction at the target site. Using this reporter system, we determined the optimal structure of SSO by investigating the effect of length, modifications, and polarities of SSOs as well as the positions of the mismatch-forming nucleotide on the efficiency of SSO-mediated gene repair. Interestingly, we found that SSOs with mismatch-forming nucleotide positioned at different positions have varying potencies that homology at the 5'-end of SSOs was more crucial for the SSO's activity. These results provided guidance for designing effective SSOs as tools for treating monogenic inherited diseases.

Increased efficiency of oligonucleotide-mediated gene repair through slowing replication fork progression.

Targeted gene modification mediated by single-stranded oligonucleotides (SSOs) holds great potential for widespread use in a number of biological and biomedical fields, including functional genomics and gene therapy. By using this approach, specific genetic changes have been created in a number of prokaryotic and eukaryotic systems. In mammalian cells, the precise mechanism of SSO-mediated chromosome alteration remains to be established, and there have been problems in obtaining reproducible targeting efficiencies. It has previously been suggested that the chromatin structure, which changes throughout the cell cycle, may be a key factor underlying these variations in efficiency. This hypothesis prompted us to systematically investigate SSO-mediated gene repair at various phases of the cell cycle in a mammalian cell line. We found that the efficiency of SSO-mediated gene repair was elevated by approximately 10-fold in thymidine-treated S-phase cells. The increase in repair frequency correlated positively with the duration of SSO/thymidine coincubation with host cells after transfection. We supply evidence suggesting that these increased repair frequencies arise from a thymidine-induced slowdown of replication fork progression. Our studies provide fresh insight into the mechanism of SSO-mediated gene repair in mammalian cells and demonstrate how its efficiency may be reliably and substantially increased.