Advanced Materials Research Vols. 998-999

Abstract: In femoral head necrosis, the cortical shell of the femoral head collapses and buckles into the cancellous bone. The purpose of this study is to explore the biomechanical characteristics of the femoral head and the necrosis region by comparing the results before and after drug treatment. In this paper, we study two patient cases with femoral head necrosis disease and establish the corresponding computational three-dimensional models. The results show that the deformation of the femur decreases slightly after the treatment, the equivalent stress distributes more evenly, and the stress magnitude reduces. The results also reveal that the volume of the necrosis in the femoral head decreases after treatment, the overall necrosis presents relatively lower equivalent stress, and the area with the relatively high equivalent stress is smaller comparing to the necrosis in femoral head before treatment.

Abstract: A method was developed for the determination of inorganic and total fluoride in Antarctic krill by using Ultrasonic Extraction and Ion Chromatography (IC). The analyte is separated by the anion exchange column AS-15 and detected with the conductive detector. The linear range of the calibration curve for fluorinion was 0.25 mg/L to 10 mg/L with a correlation coefficient of 0.9993, and the recovery of this method was 89.62% to 103.91%, and the RSD was lower than 7.09%, the LOD of this method was 0.008 mg/L and LOQ was 0.025 mg/L. Both inorganic and total fluoride in Antarctic krill were higher than GB 2762-2005, but they all much lower than ever reported.

Abstract: In this paper, using capillary zone electrophoresis method determined Vitamin C content in saposhnikovia divaricata. 20 mmol/L borax solution and 20% acetonitrile concentration, 20kV voltage, 254 nm UV detection wavelength was chosen for electrophoretic analysis. Measured vitamin C content in saposhnikovia divaricata were 0.297 mg/g (RSD=5.9%) (n=7).

Abstract: Echinomycin (Ech) is a small-molecule inhibitor of hypoxia-inducible factor-1 DNA-binding activity, which plays a crucial role in the regulation of ovarian functions in mammals. The present study was designed to test the hypothesis that hypoxia-inducible factor (HIF)-1alpha-mediated proliferation cell nuclear antigen (PCNA) expressions contributed to the follicular development in the rat ovary primed by pregnant mare serum gonadotropin (PMSG). Through the histological examination, the decrease of growing and antral follicle numbers was found after Ech treatment both in control and PMSG treated groups. And then PCNA mRNA and protein expressions were found to significantly increase in the ovaries treated with PMSG, and the similar changes were found in HIF-1alpha mRNA and protein expressions, indicating PMSG-induced follicular development may be through HIF-1alpha/PCNA signaling. Furthermore, PCNA expression was found to significantly decrease in the ovaries after Ech treatment, while HIF-1alpha mRNA and protein expression was no obviously changes. Further analysis found the changes of PCNA expression were consistent with HIF-1 activity in the ovaries, further suggesting the regulatory roles in the follicular development. Taken together, these results demonstrated this HIF-1alpha-mediated PCNA expression is one of the important mechanisms regulating the ovarian follicular development in mammals. Keywords: HIF-1alpha; PCNA; echinomycin; HIF prolyl hyodroxylase acitvity; follicular development

Abstract: The present study examined the effects of the immediate stimuli on the levels of catecholamine and single nucleotide of Chinese tree shrew by using RIA approach. When tree shrews were under immediate stimulation, compared to the control animals, the levels of the epinephrine (E), norepinephrine (NE), adenosine monophosphate (cAMP), and cyclic guanosine phosphate (cGMP) in blood were significantly increased. Also, the acute stimulation could significantly enhance the cAMP and cGMP levels of the midbrain ventral tegmental area (VTA), prefrontal cortex (PFC), hippocampus (Hipp), and testis (Testicle), and the dopamine (DA) levels in PFC, Hipp, striatum, and nucleus accumbens (NAc). However, when tree shrews were treated with reserpine, both the E and NE levels were significantly reduced (P < 0.01), indicating that the reserpine interventions could reduce or prevent those stress-related damages. Also, the mitochondrial index (the volume density, surface density and density) in hippocampus cells were significantly increased by 55.21%, 55.21% and 55.21% respectively (P<0.01), compared to those of the control animals housed. Our results demonstrated that the immediate stimulus can significantly affect the synthesis and release of epinephrine, norepinephrine, DA, cMAP, and cGMP in Chinese tree shrews. This study was supported by the following Chinese funding agencies: the National Natural Science Foundation of China (31060283), the National Social Science Fund (07BSH054), the Lanzhou Social Science Fund (07-1-64), and the Academic Innovation Team Building Program Management of Gansu Institute of Political Science and Law (GZF2013CXTD003).

Abstract: Objective: To establish composite culture system of rat bone marrow mesenchymal stem cells (BMSCs) and self-assembling peptide hydrogel RADA16-І, and to investigate the effect of RADA16-І hydrogel on neural differentiation of BMSCs. Methods: BMSCs were isolated, cultivated and labeled with green fluorescent protein (GFP), then they were inoculated on glass coverslips or in RADA16-І solution to form control group and RADA16-І group respectively. The morphological changes of BMSCs induced by neural induction medium were observed, and GFAP, NeuN and Map-2 expressions of BMSCs in each group were detected with immunofluorescence. Results: The induced BMSCs presented neuron-like change, and the rates of GFAP and NF-200 positive cells in RADA16-І group were higher than that in control group (P < 0.05). Conclusion: Self-assembling peptide RADA16-І hydrogel can promote neural differentiation of BMSCs, and which may be used as scaffold material on BMSCs transplantation for treatment of nervous system diseases.

Abstract: Abstract:Aim:We aimed to determine the effects of exhaustion exercise on HO-1 expression, the function of improving the sports ability and promoting the exercise induced fatigue in mice.Methods:Breeding, adaptively exercise BDF1 mice, 6 weeks age.They were randomly divided into normal saline control group,zinc protoporphyrin pharmacological inhibition group, the cobalt protoporphyrin drug induced group, HO-1 genetically modified group, each group was used for exhaustion exercise on the treadmill, and divided into immediate group and 24h after exhaustion groups.HO-1mRNA and protein, serum CK, BUN, urine protein were measured by using RT-PCR,Western-blot method, kinetic method, two point method and coomassie brilliant blue method. Results:The results indicated that HO-1mRNA expression increased after exhaustion exercise in the salinegroup, the other three groups changed little, HO-1 protein after exhaustion exercise of the saline group and drug inhibition group was higher than pre-exercise,24h after exercise obviously declined,24h after exhaustion exercise in drug induced group was higher than pre-exercise than immediately exercise.Exhaustion exercise time of the drug induced group(383.08±65.04min) and genetically modified mice(468.25±52.41min)was longer than the pharmacological inhibition (341.08±73.03min)and saline group(468.25±52.41min),significant variations were found in them(p<0.05). CK, BUN and urine protein removal were faster in drug induced group, the pharmacological inhibition of the group was slower. CK, BUN were faster in genetically modified group, but urine protein was in a higher level. Conclusion:Exhaustion exercise can induce skeletal muscle HO-1 expression of the wild type mice, it is not obvious in the higher HO-1 expression mice. Expression of HO-1 must be within limits, which can enhance mice exercise ability and promote the recovery of exercise fatigue.

Abstract: A DNA sequence encoding for the human proinsulin was designed according to the codon bias of Escherichia coli and then chemically synthesized. The synthesized DNA fragment was subcloned into pGEX-3X for expression in E. coli BL21 (DE3) and E. coli BL21 Star (DE3), respectively. Conditions for the highest expression of the GST-proinsulin fusion proteins were optimized. These conditions are that cells of E. coli BL21 star (DE3) are incubated in 100mL of the LB medium with 2 mmol/L IPTG and 60μg/mL ampicillin at 26^oCfor 4h. After disrupted E. coli cells with ultrasonication, inclusion bodies were precipitated from cell lysis and washed. Fusion proteins from the inclusion bodies were redissolved in 8mmol/L of urea. After dialysed in purified water, fusion proteins were analysed by SDS-PAGE. The purity of the fusion protein is about 80.5% in total. The fusion protein from SDS-PAGE was further identified by mass/mass spectrum. GST in the dyad protein is confirmed by the 9 matched sequences. However, the left part is proved a polypeptide of which is completely different from the human proinsulin.

Abstract: In order to explore the related mechanism of Foxp3 in tumor immune escape, the study detected the expression of Foxp3 in lewis lung cancer (LLC) cells and analyzed the expression of TGF-β1 and IL-10 in Foxp3 overexpressed LLC cells. Foxp3 mRNA was detected in LLC cells by RT-PCR. Foxp3 was highly expressed in Foxp3 transfected LLC group than that of in empty vector group and LLC group by RT-PCR(P <0.01). The mRNA and protein expression of TGF-β1 and IL-10 significantly increased in Foxp3 transfected LLC group than that of in empty vector group and LLC group by RT-PCR and ELISA(P <0.05). These results suggest that Foxp3 in LLC cells may promote tumor immune escape by enhancing the expression of TGF-β1 and IL-10.

Abstract: Silver nanoclusters synthesized using aptamer as matrices attracted more attention in the fields of medicine, pharmacology and biology. In this paper, we report a novel approach to sgc-8-C8-templated formation of fluorescent Ag nanoclusters and its application to bioimaging. This was further confirmed by flow cytometry. The cell toxicity (3-[4, 5-dimethylthiazolyl-2]-2, 5 -diphenyltetrazolium bromide, MTT) assay demonstrated that the silver nanoclusters has only little affect on the cytotoxicity to the cells. These results demonstrated that this aptamer-based method for labeling and imaging cellular protein is facile, effective.