In our previous work, we found that trivalent dimethylarsinous acid (DMA(III)) have high affinity binding to cysteine residue 13 of rat hemoglobin. However, it is still unknown why arsenic intermediate metabolite DMA(III) has high binding affinity for Cysl3 but not for other cysteine residues 93, 140, 111 and 125. In order to better understand the molecular mechanism of DMA(III) with rat hemoglobin, we have done current study. So, SD rats were divided into control and arsenic-treated groups randomly. Arsenic species in lysate of red blood cells were analyzed by HPLC-ICP-MS, and then determined by a hybrid quadrupole TOF MS. In addition, trivalent DMA(III) binds to different cysteine residues in rat hemoglobin alpha and beta chains were also simulated by Molecular Docking. Only Cys13 in alpha chain is able to bind to DMA(III) from the experiment results. Cys13 of alpha chain in rat hemoglobin is a specific binding site for DMA(III), and we found that amino acids compose pockets structure and surround Cys13 (but not other cysteine residues), make DMA(III) much easy to bind cysteine 13. Taken together, the DMA(III) specific binding to Cys13 is related to spatial structure of Cys13.

BACKGROUND:Previous studies have shown that composite scaffold of chitosan and poly-L-lactic acid has good biocompatibility with some cells. OBJECTIVE:To study the biocompatibility of poly-L-lactic acid reinforced by chitosan and olfactory ensheathing cells. METHODS:In experimental group, olfactory ensheathing cells from Sprague-Dawley rats aged 1-3 days were incubated onto chitosan-reinforced poly-L-lactic acid film. And in control group, olfactory ensheathing cells were co-cultured with poly-L-lysine. The proliferative ability of olfactory ensheathing cells was detected and the cells were observed with immunofluorescence histochemical staining at 1, 3, 5, 7 days after culture. RESULTS AND CONCLUSION:Olfactory ensheathing cells could survive on the chitosan-reinforced poly-L-lactic acid film, and the cytotoxic grade wasⅠ. Morphology of the cells in the experimental group was round or oval, with little processes and the cells aggregated into groups. One day after implantation, the periphery cells of the mass extended short projections and gradual y spread outward;3 days after implantation, the cells spread and most of the cells generated projections, most of which were bipolar or tri-polar;5 days after implantation, cel processes significantly extended, most cells were bipolar and tri-polar cells, while some were oval cells and irregular triangular cells;7 days after implantation, the cel density increased, and cel processes extended. Cel morphology of the control group had similar characteristics as the experimental group. There was no obvious difference between the control and the experimental group in number, perimeter or area of the cells (P>0.05). It showed that chitosan-reinforced poly-L-lactic acid had good biocompatibility with olfactory ensheathing cells.

BACKGROUND: Fibrin glue has been demonstrated to function as a kind of biomaterial with high quality. It has been used in nerve tissue engineering and proved to be a kind of scaffold for some cells.OBJECTIVE: To explore the biocompatibility of fibrin glue and olfactory ensheathing cells (OECs).DESIGN, TIME AND SETTING: An in vitro control trial based on cytology was performed at the Institute of Neurobiology,Nantong University from August 2007 to February 2008.MATERIALS: Fibrin glue was made of fibrin and catalyst, and OECs derived from rats' olfactory bulb were normally primary-cultured.METHODS: OECs were divided into control (OECs clone spheres were cultured alone) and in fibrin glue (OECs clone spheres were cultured and combined with fibrin glue) groups. After 1 week of culture, the proliferation of OECs were observed by convert microscope and detected by S-100 immunofluorescence histochemical staining.MAIN OUTCOME M EASURES: OECs morphology, cell count, the area of the cell bodies and the perimeter of the cell were determined.RESULTS: OECs could survive, migrate in fibrin glue, and float in the fibrin glue in the lower layer. After 7 days of incubation, cell body exhibited fusiform or triangle, predominantly bipolar or bipolar. The number of the S-100 positive cells was more, and cell bodies were larger in fibrin glue group than control group (P < 0.05). However, there was no obvious difference between two groups in cell perimeter (P > 0.05).CONCLUSION: Fibrin glue has good biocompatibility with OECs, and OECs can survive and migrate in fibrin glue.

Objective To observe the efficacy of combination therapy of calcium dobesilate dispersible and monosialotetrahexosylganlioside sodium on interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in elderly patients with painful diabetic peripheral neuropathy.Methods From January 2012 to May 2017,in the Second People's Hospital of Lianyungang 70 patients of painful diabetic peripheral neuropathy,aged ≥60 years,were analyzed in this study.They were randomly divided into observation group (35 cases) and control group (35 cases).The observation group was treated with 40mg monosialotetrahexosylganlioside sodium dissolved in 250mL physiological saline,intravenous infusion per day,and oral calcium dobesilate dispersible 0.5g twice a day for two weeks.The control group was treated with methylcobalamin injection 0.5mg per day for two weeks.The clinical treatment effects and levels of IL-6 and MCP-1 were observed and compared between the two groups.Results After two weeks of treatment,the MDNS and MNSI scores of the observation group [(13.09 ± 5.38)points,(2.53 ± 1.19)points] were significantly lower than those of the control group [(18.31 ± 6.13) points,(4.19 ± 1.05) points,t =2.036,2.365,all P ＜ 0.05] and those before treatment [(21.26 ± 4.28) points,(5.40 ± 0.89) points,t =3.251,3.698,all P ＜ 0.05].The VAS-PI scores in the observation group [(6.24 ± 1.25) points,(4.13 ± 1.69) points] were significantly lower than those in the control group[(7.26 ± 1.28) points,(6.34 ± 2.65) points] at the first and second week (t =3.265,5.395,all P ＜ 0.05).The serum levels of inflammatory cytokines IL-6 and MCP-1 in the observation group [(15.16 ±0.88) ng/L,(157.19 ± 11.22) ng,/L] were significantly lower than those in the control group[(17.87 ± 1.19) ng/L,(198.21 ± 12.07)ng/L,t =2.152,1.365,all P ＜0.05]and those before treatment[(20.26 ± 1.05) ng/L,(260.44 ± 13.63) ng,/L,t =1.235,0.965,all P ＜ 0.05].Conclusion Combination of calcium dobesilate and mono-sialotetrahexosyl ganglioside may alleviate the sensory and pain sensations in patients with painful diabetic peripheral neuropathy,possibly by reducing the level of inflammatory cytokines IL-6 and MCP-1.

Objective To investigate the effects of duration of surgery on flash-induced visual evoked potentials (VEP) in patients undergoing spinal surgery in prone position.Methods Eighty-two ASA Ⅰ or Ⅱ patients of both sexes aged 20-76 yr weighing 43-96 kg undergoing spinal surgery in prone position were divided into 3 groups according to the duration of surgery:group S≤2 h ( n =34) ; group M 2-4 h ( n =38) and group L≥4 h ( n =10).VEP was monitored using protektor VEP monitoring device (Xltek Co.,Canada).The latency,amplitude and recovery time of wave P100 were recorded before and 10 min after induction of anesthesia and at the end of surgery.Results Compared with group S,the amplitude of wave P1000 was significantly decreased at the end of surgery in group M,the lantency of wave P100 was significantly prolonged,while the amplitude of wave P100 was decreased at the end of surgery in group L ( P ＜ 0.05).Compared with group M,the lantency of wave P100 was significantly prolonged,while the amplitude of wave P100 was decreased at the end of surgery in group L ( P ＜ 0.05).Compared with groups S and M,the recovery time of wave P100 was significantly prolonged in group L ( P ＜0.05).There was no significant difference in the recovery time of wave P100 between groups S and M ( P ＞ 0.05).Conclusion Duration of surgery (≥4 h) can affect flash-induced VEP,the longer the duration,the stronger the effects.

The present research was aimed to explore the biocompatibility of IKVAV self-assembling peptide nanofiber scaffold with olfactory ensheathing cells (OECs) of rats. The OECs were seeded onto the surface of coverslips covered with IKVAV self-assembling peptide nanofiber scaffold hydrogel (2D culture system), and implanted within IKVAV self-assembling peptide nanofiber scaffold hydrogel (3D culture system), respectively. The adhesion, viability of OECs were observed with inverted microscope. Then the characteristics for survival and adhesion of cells by image processing were observed, and statistical analysis on the number of S-100 positive cell, the area of the cell bodies and the perimeter of the cell and MTT method were carried out. It was found that the OECs could survive and migrate in IKVAV self-assembling peptide nanofiber scaffold. The result of the cell MTT exam, of the shape and quantity of cells had no significant difference compared to those of the OECs cultured with poly-L-lysine (PLL). It has been proved that IKVAV self-assembling peptide nanofiber scaffold has good biocompatibility with rat OECs.
Animals ; Animals, Newborn ; Biocompatible Materials ; chemistry ; Cell Proliferation ; Cells, Cultured ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Laminin ; chemistry ; Nanofibers ; chemistry ; Olfactory Bulb ; cytology ; drug effects ; Peptide Fragments ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

Objective:To analyze the effect of panax notoginseng saponins(PNS)on mTORC1-HIF1α signaling pathway,and to explore its effect and mechanisms on the differentiation balance of Th17/Treg cells in CD4+T cells.Methods:Isolate the spleens of C57BL/6 mice,then select CD4+T cells by magnetic beads and cultured in vitro.The optimal concentration of PNS was screened by the CCK-8,and then these cells were divided into control group and PNS treatment group(5,10 and 20 μg/ml),each gives correspond-ing drug treatment after 48 h.Afterwards,flow cytometry was used to detect differentiation of Th17/Treg cells.Real-time quantitative fluorescent PCR was used to detect the expressions of RORγt,Foxp3,mTOR,Raptor,HIF1α mRNA.ELISA was used to detect the levels of IL-17A and IL-10 in the supernatant of cell culture.Western blot was used to detect the expressions and phosphorylation levels of 4EBP1,S6K and HIF1α proteins.Results:5,10,20 μg/ml PNS could significantly inhibit Th17 cells differentiation and promote Treg cells differentiation;5,10,20 μg/ml PNS could significantly reduce the expression of RORγt mRNA,and then reduce the level of IL-17A;20 μg/ml PNS could significantly promote the expression of Foxp3 mRNA and increase the level of IL-10;10,20 μg/ml PNS could significantly decrease the phosphorylation of 4EBP1 and S6K;5,10,20 μg/ml PNS could significantly reduce the expression of HIF1α mRNA and inhibit the expression of HIF1α protein.Conclusion:Certain concentrations of PNS can inhibit the differentiation of Th17 cells in CD4+T cells,and promote the differentiation of Treg cells,which is related with modulating mTORC1-HIF1α signaling pathway.

Objective To investigate the relationship between plasma tau protein, phosphorylated tau protein (p-tau) protein and cognitive function in subjects with generalized brain atrophy. Methods A total of 100 subjects with moderate and severe brain atrophy were divided into two groups according to cognitive function: normal group (n=50 cases) and dementia group (n=50 cases). And their gender, age, educational level, etc. are recorded. The tau protein and p-tau protein content in plasma were measured by double antibody sandwich enzyme-linked immunosorbent assay (ELISA). The differences between plasm tau and p-tau protein expression and their relationship with cognitive function were analyzed. Results Plasma tau protein and p-tau protein levels were significantly higher (P<0.05) in the dementia group [(210.92±43.79)pg/mL、(81.15±16.85)pg/mL] than in the normal group[(210.92±43.79)pg/mL、(81.15±16.85)pg/mL]. Plasma tau protein and p-tau protein levels were negatively correlated with the MMSE score (P<0.05) and had no significant correlation with the degree of brain atrophy (P>0.05). Conclusion Cognitive impairment may be associated with elevated tau protein levels in patients with extensive brain atrophy.

Objective:To investigate the effects of poly adenosine diphosphate ribose polymerase-1(PARP-1) inhibitor fluzoparib on proliferation, apoptosis and migration of pancreatic cancer PANC1 cells.Methods:PANC1 cells cultured in conventional culture medium were used as control group, and PANC1 cells cultured in the medium containing fluzoparib were used as fluzoparib group. The effects of fluzoparib with different concentrations on the proliferation of PANC1 cells were detected by CCK8 method, and the half inhibitory concentration (IC 50) of fluzoparib on PANC1 cells was calculated. The effect of fluzoparib on apoptosis and cell cycle of PANC1 cells was detected by flow cytometry, and the migration ability of PANC1 cells was detected by cell scratch test and Transwell chamber. Results:Compared with control group, with the increase of fluzoparib concentration and the prolongation of the action time, the cell proliferation activity of PANC1 in fluzoparib group was significantly decreased, and the differences were statistically significant (all P values <0.05). IC 50 of fluzoparib on PANC1 cells cultured for 24 h was 0.03 mmol/L. After 24 h culture, the IC 50 apoptosis rate of fluzoparib group was (32.19±2.48)%, and the apoptosis rate of control group was (21.99±6.30)%. The former was greatly higher than the latter, and the difference was statistically significant ( P<0.05). The proportion of cells in G 2/M phase was (16.28±0.62)% in the fluzoparib group and (11.64±0.88)% in the control group, and the difference between the two groups was statistically significant ( P<0.05). The migration rates of PANC1 cells in IC 50 fluzoparib group in 12 h and 24 h culture were (2.59±1.46)% and (19.76±7.84)%; and those in control group were (27.08±2.17)% and (45.92±3.61)%, respectively. The number of transmembrane cells was (348±19) cells/10 visual field in the fluzoparib group and (587±14) cells/10 visual field in the control group. The migration ability of PANC1 cells in fluzoparib group was significantly lower than that in control group ( P<0.05). Conclusions:Fluzoparib can inhibit the proliferation and migration of PANC1 cells and promote the apoptosis of PANC1 in vitro, which may be an effective drug for the treatment of pancreatic cancer.

Objective To explore the correlations between serum soluble growth stimulation gene 2 protein(sST2),brain natriuretic peptide(BNP)and the degree of carotid atherosclerosis in patients with acute ischemic stroke.Methods A total of 205 patients with acute ischemic stroke were selected as study group,and were divided into mild group with 100 cases,moderate group with 70 cases,and severe group with 35 cases according to the severity of carotid atherosclerosis.Another 150 healthy people who received physical examination during the same period were selected as control group.Serum levels of sST2 and BNP were compared among all groups.The correlations of sST2 and BNP levels with the degree of carotid atherosclerosis were analyzed.The influencing factors of different degree of carotid atherosclerosis in patients with acute ischemic stroke were analyzed.The predictive value of sST2 combined with BNP on the degree of carotid atherosclerosis in patients with acute is-chemic stroke was analyzed.Results The serum sST2 level of the study group was significantly lower than that of the control group,and the serum BNP level was significantly higher than that of the control group(P＜0.05).The sST2 levels in moderate and severe groups were significantly low-er,BNP levels were significantly higher than that in the mild group(P＜0.05).BNP level in the severe group was significantly higher,sST2 level was significantly lower than that in the moderate group(P＜0.05).The degree of carotid atherosclerosis in patients with acute ischemic stroke was correlated with age,history of hypertension,history of smoking,history of diabetes,levels of total cholesterol(TC),C-reactive protein(CRP),homocysteine(Hcy)and low density lipoprotein(LDL)(P＜0.05).Age,hypertension history,smoking history,diabetes history,TC,CRP,Hcy,LDL,BNP and sST2 were the influencing factors for degree of carotid atherosclerosis(P＜0.05).Serum BNP level was positively correlated with the degree of carotid atherosclerosis,while sST2 level was negatively correlated with the degree of carotid atherosclerosis(P＜0.001).The ar-ea under the curve(AUC)for the combined prediction of sST2 and BNP was significantly greater than that for the individual prediction of sST2 and BNP(P＜0.001).Conclusion Clinical obser-vation of the changes in serum levels of sST2 and BNP in patients with acute ischemic stroke can ef-fectively assess the progression of the disease,which is beneficial for improving the severity of carot-id atherosclerosis in patients.The combined assessment of sST2 and BNP shows good diagnostic per-formance for evaluating the severity of carotid atherosclerosis in patients with acute ischemic stroke.