Substance P combined with epidermal stem cells promotes wound healing and nerve regeneration in diabetes mellitus

Exogenous substance P accelerates wound healing in diabetes,but the mechanism remains poorly understood.Here,we established a rat model by intraperitoneally injecting streptozotocin.Four wounds（1.8 cm diameter） were drilled using a self-made punch onto the back,bilateral to the vertebral column,and then treated using amniotic membrane with epidermal stem cells and/or substance P around and in the middle of the wounds.With the combined treatment the wound-healing rate was 100% at 14 days.With prolonged time,type I collagen content gradually increased,yet type III collagen content gradually diminished.Abundant protein gene product 9.5-and substance P-immunoreactive nerve fibers regenerated.Partial nerve fiber endings extended to the epidermis.The therapeutic effects of combined substance P and epidermal stem cells were better than with amniotic membrane and either factor alone.Our results suggest that the combination of substance P and epidermal stem cells effectively contributes to nerve regeneration and wound healing in diabetic rats.

Microencapsulation improves inhibitory effects of transplanted olfactory ensheathing cells on pain after sciatic nerve injury

Olfactory bulb tissue transplantation inhibits P2X2/3 receptor-mediated neuropathic pain. However, the olfactory bulb has a complex cellular composition, and the mechanism underlying the action of purified transplanted olfactory ensheathing cells（OECs） remains unclear. In the present study, we microencapsulated OECs in alginic acid, and transplanted free and microencapsulated OECs into the region surrounding the injured sciatic nerve in rat models of chronic constriction injury. We assessed mechanical nociception in the rat models 7 and 14 days after surgery by measuring paw withdrawal threshold, and examined P2X2/3 receptor expression in L4–5 dorsal root ganglia using immunohistochemistry. Rats that received free and microencapsulated OEC transplants showed greater withdrawal thresholds than untreated model rats, and weaker P2X2/3 receptor immunoreactivity in dorsal root ganglia. At 14 days, paw withdrawal threshold was much higher in the microencapsulated OEC-treated animals. Our results confirm that microencapsulated OEC transplantation suppresses P2X2/3 receptor expression in L4–5 dorsal root ganglia in rat models of neuropathic pain and reduces allodynia, and also suggest that transplantation of microencapsulated OECs is more effective than transplantation of free OECs for the treatment of neuropathic pain.

Down-regulation of Halr1 during induced differentiation of embryonal carcinoma P19 cells

Maintenance of pluripotency depends to diverse regulatory factors.Studies in embryonic stem cells(ESCs)have indicated that large intergenic non-coding RNAs(lincRNAs)are involved in the regulatory network of pluripotency.However,the presence and function of pluripotency-associated lincRNAs in cancer cells with pluripotency features are unknown.In this study,we used embryonal carcinoma(EC)P19 cell lines to investigate the expression level of Halr1 in pluripotency and retinoic acid(RA)-induced differentiated states.Down-regulation of pluripotency associated factors such as OCT4,NANOG,SSEA1 and alkaline phosphatase at transcript and protein levels were used to confirm the differentiated status of P19 cells.Quantitative measurement of Halr1 transcript levels revealed a 79% decrease during RA-induced differentiation of P19 cells.These results indicate that upon exiting the pluripotency state the expression level of Halr1 similar to core pluripotency factors is remarkably reduced.

circ_0003204 regulates the osteogenic differentiation of human adipose-derived stem cells via miR-370-3p/HDAC4 axis

Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in mesenchymal stem cell osteogenesis.However,how circRNAs regulate hASCs in osteogenesis is still unclear.Herein,we found circ_0003204 was significantly downregulated during osteogenic differentiation of hASCs.Knockdown of circ_0003204 by si RNA or overexpression by lentivirus confirmed circ_0003204 could negatively regulate the osteogenic differentiation of hASCs.We performed dual-luciferase reporting assay and rescue experiments to verify circ_0003204 regulated osteogenic differentiation via sponging miR-370-3p.We predicted and confirmed that miR-370-3p had targets in the 3′-UTR of HDAC4 m RNA.The following rescue experiments indicated that circ_0003204 regulated the osteogenic differentiation of hASCs via miR-370-3p/HDAC4 axis.Subsequent in vivo experiments showed the silencing of circ_0003204 increased the bone formation and promoted the expression of osteogenic-related proteins in a mouse bone defect model,while overexpression of circ_0003204 inhibited bone defect repair.Our findings indicated that circ_0003204 might be a promising target to promote the efficacy of hASCs in repairing bone defects.

Changes of the expression of CCNL2 gene during the differentiation of P19 cells to cardiac myocytes

Objective： To investigate the changes of cyclin L2（CCNL2） gene mRNA and protein during the differentiation of P19 cells to cardiac myocytes, and to explore the relationship between CCNL2 gene and the differentiation of cardiac myocytes. Methods： P19 cells were cultured with 0.9% DMSO for 18 days. Western blots of cardiac troponin I （cTnI） were used to identify cell differentiation. Total RNA was extracted from P19 cells during the process of differentiation at various time points：pre-differentiation（Day 0）, and Day 1 to Day 18. The expression levels of CCNL2 gene mRNA and protein were evaluated by RT-PCR and Western blot, respectively. Results： After being induced to differentiate by DMSO for 4 days in suspension, spontaneously and rhythmically beating ceils were seen at 8 day, which were cTnI-positive. In P19 cells, both the expression level of CCNL2 gene mRNA and protein were gradually down-regulated. Conclusion： Both the expression of CCNL2 gene and protein were down-regulated during the process of the differentiation of P19 cells into cardiac myocytes, suggesting a possible role for this cyclin in their differentiation.

Effect and mechanism of high glucose on apoptosis and cell cycle arrest of islet β cells via p27 pathway

Objective:To study the effect of mechanism of high glucose on apoptosis and cell cycle arrest of isletβcells via p27 pathway.Methods:Islet INS-1 cells were cultured and divided into groups.The control group was treated with ordinary medium,the high glucose group was treated with high glucose medium containing 25mmol/L glucose,the high glucose+si-NC group was treated with high glucose medium and transfected with NC siRNA,and the high glucose+si-P27 group was treated with high glucose medium and transfected with p27 siRNA.After 24 hours treatment,MTS assay was used to detect the cell viability A490,TUNEL assay was used to detect apoptosis rate,flow cytometry was used to detect the cell cycle distribution and western blot was used to detect the expression levels of P27,caspase-8 and cyclinD1.Results:Compared with those in the control group,the A490,the ratio of S phase and G2/M phase as well as the expression level of CyclinD1 decreased,while the apoptosis rate,the ratio of G0/G1 phase as well as the expression levels of P27 and caspase-8 increased in the high glucose group(P<0.05);compared with those in the high glucose group,the A490,cell cycle as well as the expression levels of P27,caspase-8 and cyclinD1 were not different from those in the high glucose+si-NC group(P>0.05);compared with those in the high glucose group and high glucose+si-NC group,the A490,the ratio of S phase and G2/M phase as well as the expression levels of cyclinD1 increased,while the apoptosis rate,the ratio of G0/G1 phase as well as the expression levels of p27 and caspase-8 decreased in the high glucose+si-P27 group(P<0.05).Conclusion:The apoptosis and cell cycle arrest induced by high glucose are related to P27 pathway activation.

P2X receptors in maintenance and differentiation of neural progenitor cells

Purinergic receptors are among the first cell surface receptors expressed during embryonic development （Burnstock and Ulrich, 2011）. These are characterized based on their pharmacological properties of being activated by adenosine or purine/pyrimidine nucleotides as P1 and P2 receptors. P2 receptors are further classified by their structure as P2Y metabotropic and P2X ionotropic receptors.

Targeting the P2X7 receptor in microglial cells to prevent brain inflammation

Microglial cells are the key innate immune cells in the brain and they are crucial in maintaining brain parenchyma homeostasis.Under physiological conditions,microglial cells assume a ramified morphology with a small cell body and an extensive network of fine processes,which secrete neurotrophic factors and patrol the surroundings in search for pathogens and eliminate cellular debris via phagocytosis.Microglial cells express a repertoire of pattern recognition receptors(PRRs)that enable them to detect diverse danger-associated molecular patterns(DAMPs)released from damaged cells or cells under stress,or pathogen-associated molecular patterns generated by pathogens during infection.

p53 and NFκB regulate microRNA-34c expression in porcine ovarian granulosa cells

supported by the National Natural Science Foundation of China （31201771）;the earmarked fund for the China Agriculture Research System （CARS-36）

A Creation Model from the Gell-Mann Standard Model to the Creation of Bio Cells: Based on the Assumption of Homogeneous 5D Space-Time Universe

In this paper, we briefly go over the homogeneous 5D model field theory: from the 5D space-time inception, to its quantum field solutions given in terms of Higgs vacuum, filled with magnetic monopole bose fields of all energies. Then through the space dimension reduction projections, the Gell-Mann standard model was obtained as well as a quantum to Classical connection was made via introducing Bose distribution to the monopoles to obtain the Perelman entropy and Ricci Flow mappings. This provided us a picture to the creation of Astronomical objects, from galaxies to stars and planets. This method of splitting the monopole energy into ranges is extended to show that below the basic rest mass range of the electron and Quark, it still can be applied to explaining for the creation of the chemical elements periodic table. But perhaps the most interesting is in the lowest hundreds of Hz energy range, obtained from yet another 3 fold space symmetry breaking, into 2D × 1D, producing bio nitrogenous bases composed of 3 Carbon 12 in hexagon structures, due to preservation of the 1D monopole standing waves of this low frequencies. From that by imposing gauge changes the monopole states into DNA spectra. Since such spectra states retain the DLRO, it induces formation of charge carriers periodicity in a spherical bio cell.. It was then argued that due to cell’s surface proteins, the structure must contain partial filled VB, with “p” state hole density, and empty CB, separated from VB by a positive band gap. Such band structures resemble known HTC Cuprate ceramics. Since the HTC goes through a Superconductivity transition via the simultaneous bose exciton condensation, providing a Coulomb pressure, which reduces the band gap substantially, and induces the ODLRO transition of the hole density. The same obviously applies to the bio cells. Because of the near continuous exciton levels generated, a matching to the DNA spectra then can always occur by selective choices of proteins on the cell surface. Judging from a numerical study, we did years ago on YBCO, with doping. We found with a large enough VB hole density, the exciton induced superconducting gap can easily lead to Tc in the room temperature range. In fact by EMF excitation can increase the exciton pressure and trigger the ODLRO transition Tc upward. In fact, numerical results then suggest there do exist coherent EMF spectra from three key elements: Water, Carbon and Hydrogen, together with Oxygen, as studied over the years by numerous people, starting from Schrödinger to most recently Geesink.

Effect of Valence Band Tail Width on the Open Circuit Voltage of P3HT:PCBM Bulk Heterojunction Solar Cell:AMPS-1D Simulation Study

The effect of the valence band tail width on the open circuit voltage of P3HT：PCBM bulk heterojunction solar cell is investigated by using the AMPS-1D computer program. An effective medium model with exponential valence and conduction band tail states is used to simulate the photovoltaic cell. The simulation result shows that the open circuit voltage depends Iinearly on the logarithm of the generation rate and the slope depends on the width of the valence band tail. The open circuit voltage decreases with the increasing width of the band tail. The dark and light ideality factors increase with the width of the valence band tail.

Regulation of neural stem/progenitor cell functions by P2X and P2Y receptors

Neural stem/progenitor cells：Radial glial cells constitute multipotent cells in the ventricular zone,lining the wall of the lateral ventricle of the embryonic brain.They have the capacity to give rise to cells belonging to all three major linages（neurons,astrocytes and oligodendrocytes）of the nervous system（Tang and Illes,2017）.

Expression of mTOR/P70 S6K in oral acinic cell carcinoma

This study was aimed to observe the expression of P70 S6 kinase （P70 S6K） in oral acinic cell carcinoma. P70 S6 kinase expression was examined by means of Western-blot test and Activity assay. Specimens were from 30 cases of oral acinic cell carcinoma and 15 cases of normal oral tissue were used as controls. Statistical analysis software SPSS10.0 was used for t test to determine the relationship between gene expression and clinical features. The expression level of P70 S6K increased obviously in oral acinic cell carcinoma tissue （ P 〈 0.01 ）. Activity assay was the same as the Westemblot test （ P 〈 0.01 ）. P70 S6K expression level and activity played an important role in the development of oral acinic cell carcinoma. In conclusion, P70 S6K is amplified and overexpressed in oral acinic cell carcinoma tissue, which suggests a potential oncogenic function. P70 S6K and other possible targets of roTOR contribute significantly to tumor development and that inhibition of these proteins may be therapeutic for cancer patients. Overexpression of P70 S6K may be involved in the pathogenesis of oral acinic cell carcinoma.

Observations on the Contraction and Movement of Fat-Storing Cells in Culture

The Wister fat-storing cells were isolated by perfusion with collagenase and centrifugation with metrizamide density cushion technique and cultured in vitro. The fatstoring cells were confirmed by the presentation of the lipid drop in the cytoplasm and the visualization of dismin with anti-dismin antibody by using indirect immunofluorescence method. By the videotape recorder (VIR) and the imagine analysis system, we observed the wandering immigration, the abrupt contraction of fat-storing cells with spike, then becoming a ball-like shape in its division phase. After that the cell began to extend and the contraction of these cells can be induced by the presence of 10-2 mmol/L endothelin-1 , 1 mmol/L of substance P and 2×10-5 mmol/L noradrenalin. After removal of these agents the contracted cells would become extended. All these findings indicate that the fat-storing cells have ability of contraction and movement.

Retroviral vector containing human p16 gene and its inhibitory effect on Bcap-37 breast cancer cells

Objective To study the effects of the p16 gene on tumor cell growth inhibition and cell cycle arrest. Methods The recombinant retroviral vector pLp16SN was constructed by cloning the human p16 cDNA into the retroviral vector pLXSN. Retroviruses with or without the p16 gene were obtained by transfecting pLp16SN and pLXSN vectors into PA317 cells. Bcap-37 human breast cancer cells were infected with these retroviruses followed by selection with G418. The expression of p16 was detected by Northern and Westem blots. Cell biological characteristics, including cell growth rate, cell cycle and tumorigenesis in nude mice were assessed.Results Both mRNA and protein expression of p16 in Bcap-37 cells transfected with retroviral vector containing the p16 gene were much higher than that in Bcap-37 cells transfected with empty vector or parental Bcap-37 cells. Cell overexpressing the p16 gene exhibited a slower rate of growth, a higher percentage of cells in the G1 phase, and smaller tumors in nude mice, compared with parental Bcap-37 cells and cells transfected with empty vector.Conclusion Overexpression of the p16 gene could suppress the growth of Bcap-37 breast cancer cells by arresting the cell cvcle at the G1 to S-phase.

InGaAsP/InGaAs tandem photovoltaic devices for four-junction solar cells

Lattice-matched lnGaAs（P） photovoltaic devices were grown on lnP substrates by metal-organic chem- ical vapor deposition. InGaAsP/InGaAs （ 1.07/0.74 eV） dual-junction （D J） solar cells were fabricated and charac- terized by quantum efficiency and I-V measurements. The open circuit voltage, short circuit current density, fill factor, and efficiency of lnGaAsP/lnGaAs DJ solar cell are 0.977 V, 10.2 mA/cm2, 80.8%, and 8.94%, respectively, under one sun illumination of the AM 1.5D spectrum. For the lnGaAsP/lnGaAs DJ solar cell, with increasing con- centration, the conversion efficiency first increases steadily and reaches 13% around 280 suns, and finally decreases due to the drop in fill factor at higher concentration ratios. These experimental results demonstrate the promising prospect of GaInP/GaAs/lnGaAsP/lnGaAs four-junction solar cells.

Construction of antisense Bmi-1 expression plasmid and its inhibitory effect on K562 cells proliferation

Background Bmi-1 gene determines the proliferative capacity of normal and leukemia stem cells. Expression of Bmi-1 has been found in all types of myeloid leukemia cells in both humans and mice. This study aimed at assessing the effect of antisense Bmi-1 expression on K562 cells proliferation and p16 protein （p16） expression. Methods A transcriptional repressor, Bmi-1 cDNA was cloned by reverse transcriptase polymerase chain reaction （RT-PCR） of its mRNA from K562 cells. A plasmid expressing antisense Bmi-1 mRNA was then constructed by reverse design of PCR primers and cloned to the plasmid pLNCX2; G418 was added to the medium after the plasmid was successfully introduced in K562 cells by lipofectin-mediated DNA transfection. The effects of the antisense expression on the proliferation of K562 cells were analyzed by using microcuhure tetrazolium and colony forming. Cell cycle was analyzed by using flow cytometry. The p16 expression of K562 cells was observed by immunoflorescence histochemical stain. Results K562 cells transfected with antisense Bmi-1 plasmid grew significantly slower than that of controls （the parental K562 and cells transfected with empty plasmid）. The colony forming ability of antisense Bmi-1 plasmid transfected ceils decreased significantly （P 〈0. O1 ） compared with controls. The p16 expression of cells transfected with antisense Bmi-1 was upgraded more apparently than that of controls. Conclusion The antisense Bmi-1 gene can inhibit the growth of K562 cell and upgrade expression of p16 in K562 cells.

Bulk heterojunction thin film formation by single and dual feed ultrasonic spray method for application in organic solar cells

We here present a way of preparing the polymer： fullerene BHJ using dual feed method which can lead to formation of pure phases. In this report, we present results of our initial experiments in this direction.The effect of process parameters on the thickness and surface roughness of the active layer has been discussed.The structural and optical properties have been studied using the optical microscope, UV–visible spectroscopy and photoluminescence spectroscopy. Significant PL quenching indicates efficient charge separation in the BHJ formed using this technique. We have also compared the BHJ thin films prepared with this dual feed ultrasonic technique with the single feed spray method. The BHJ formed using this technique has been used as an active layer in OSC.

In Vitro and In Silico Studies of Lunacridine from Lunasia Amara Blanco as Anticancer

Lunasia amara Blanco is a famous plant in South Sulawesi. It was used largely by local people as antibacteria and aphrodisiac. Quinoline alkaloid lunacridine was known as the active principle from Lunasia amara Blanco. Its activity was reported as a DNA Intercalating Topoisomerase II inhibitor. In this study, we have isolated and assayed the cytotoxic activity of lunacridine on P388 murine leukemia cells by MTT colorimetric assay （in vitro）. Lunacridine showed the less cytotoxic activity with the IC50 of 39.52 μg/mL. With the aim to explore the structural determinants responsible for this activity, molecular docking study have been carried out with DNA model using AutoDock 4.0 software with various total of energy evaluations （in silico）. The best docking reached at the total energy evaluations of 2.5 × 107 with the binding free energy of-6.63 kcal/mol. Analysis of the docking results was in accordance with the ability of lunacridine to intercalate between base pairs of DNA. Cytotoxic activity of lunacridine is less probably due to low affinity and molecular interaction. Therefore this study suggests to design and to develop lunacridine as a lead compound for anticancer drug.

Effects of human microRNA-181a-5p on proliferation and migration of gastric cancer cells

Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal high metastasis gastric cancer cell line GC9811-P were tested by quantitative real-time polymerase chain reaction(qRT-PCR).GC9811 cell line was