In this paper, the effects of the blend of nanoparticles and microneedle matrix materials on the mechanical properties of dissolving microneedles were studied mainly, so as to construct microneedles with excellent mechanical properties. Different kinds of nanoparticles (calcium carbonate, hydroxyapatite, silica), particle sizes (20, 60, 100 nm) and the proportion of prescription (2%, 6%, 10%) were blended with the matrix material [polyvinyl pyrrolidone (PVP), poly(1-vinylpyrrolidone-co-vinyl acetate)(PVP/VA)] to form dissolving microneedles. The effects of nanoparticles on the elastic modulus and hardness of the microneedles were investigated using a nanoindenter. The results showed that the elastic modulus and hardness of PVP microneedles were significantly improved by nano-calcium carbonate (P < 0.001), and the elastic modulus and hardness of PVP/VA microneedles were significantly improved by nano-hydroxyapatite (P < 0.001). When the particle size of hydroxyapatite was 20 nm, the elastic modulus of PVP/VA microneedles was (10.6 ± 1.0) GPa, and the hardness was (0.47 ± 0.06) GPa. As the size of the nanoparticles increases, the mechanical performance of the microneedles decreases. When the mass proportion of nano-hydroxyapatite increased from 2% to 6%, the elastic modulus and hardness of the microneedles were significantly improved (P < 0.001), but the effect of continue increasing the proportion of nanoparticles on the microneedles was not significant. The nano-enhanced PVP/VA dissolving microneedles has no irritant effect on intact skin and has a slight irritation to damaged skin, but they disappear completely after 72 h. Animal experiments have been approved by the Laboratory Animal Welfare and Ethics Committee of Zhejiang University of Technology. Therefore, the nano-enhanced dissolving microneedles has good biological safety. To sum up, it is necessary to select the appropriate kind of nanoparticle, particle size, and prescription ratio when microneedles constructing with a given matrix material, so as to effectively improve its mechanical performance.

Humans ; Nasal Cavity ; Paranasal Sinuses

Objective To investigate the effects of human Rrain-derived neurotrophic factor-gamma fetopro-tein (hBDNF-GFP) gene-transfected neural stem cell (NSC) transplantation on BDNF expressions in the retina of rots after optic nerve (ON) crush injury. Method ①Seventy-eight Sprague-Dawley (SD) rats were randomly as-signed into a control group (n = 6) and ON crush group (n = 72). In the ON crush group, the right ON was crushed while the left NO was exposed as sham injury. Rats in the ON crush group were divided into three sub-groups: PBS group (intravitreons injection of 0.01 mol/L phosphate buffered solution); GFP group (intravitreous transplantation of GFP gene-transfected NSCs); and hBDNF-GFP group (intravitreous transplantation of hBDNF-GFP gene-transfected NSCs). Rats were sacrificed 3, 7, 14 and 28 days after transplantation, and BDNF expres-sions in retinal homogenates was detected by using enzyme-linked immunosorbent assay (ELISA). ②The hBDNF-GFP-NSCs were transplanted intravitreous into six rats after ON crush injury. Following this, two rats were sacri-riced 2, 4 and 8 weeks after transplantation. The survival and location of NSCs in host retina were observed by frozen section analysis. ③Adult SD rats were randomly divided into four groups: control group (n = 5); NSC group (NSC transplantation, n = 10); GFP-NSC group (GFP-NSC transplantation, n = 10); and hBDNF-GFP-NSC group (hBDNF-GFP-NSC transplantation, n = 10). Four and eight weeks after transplantation, five rats from every group were sacrificed. Western blot analysis was used to determine retinal BDNF expression. Results ① There was no significant difference in BDNF expression between the control group and sham-injury groups (P >0.05). Three days after NSC transplantation, BDNF expression increased significantly in the three injured sub-groups compared with the sham-injury group, (P < 0.05), whereas no significant inter-group differences in BDNF expressions among three injured sub-groups were observed (P > 0.05). Seven days after transplantation, there was a significant difference in BDNF expression between the GFP-NSC group and the sham-injury groups (P <0.05), whereas there were no significant differences in BDNF expressions among the PBS, hBDNF-GFP-NSC and sham-injury groups (P > 0.05). Fourteen and 28 days after transplantation, BDNF expressions decreased in the PBS group and the GFP-NSC groups, while BDNF expressions in the hBDNF-GFP-NSC group increased significant-ly compared with the other three groups (P < 0.05); ②Frozen section showed that transplanted hBDNF-GFP-NSCs could survive and gradually extended to all layers of the host retina. ③Westem blot revealed there were no differences in BDNF expressions between 4-week and 8-week intervals in the hBDNF-GFP-NSC group. Compared with other three groups, BDNF expressions in the retina increased significantly after hBDNF-GFP-NSC transplanta-tion. Conclusions The hBDNF-GFP gene-trausfected NSCs can survive in the host retina and BDNF expressions are stable at a high level.

Isoprene is an important precursor of synthetic rubber material. In our previous study, metabolic engineered Escherichia coli strain (BW-01) was constructed and used to produce isoprene. Based on the theory of protein budget, using synthetic biology strategies including the increased copy number of genes and rare codons, we regulated the expression of key enzyme to improve isoprene production in Escherichia coli strain. Under shake-flask conditions, isoprene productivity of the engineered strain (BW-07) increased by 73% compared with BW-01, reached 761.1 mg/L. It provides a reference for further studies.
Butadienes ; Escherichia coli ; genetics ; metabolism ; Gene Dosage ; Hemiterpenes ; biosynthesis ; Industrial Microbiology ; Metabolic Engineering ; Mevalonic Acid ; Pentanes ; Synthetic Biology

BACKGROUND: Myelin sheath related inhibitors have been found to have great impact on microenvironment of axon regeneration. Traditional Chinese medicine is gradual y becoming a research hotspot on improving microenvironment of nerve regeneration with its advantage on multiple factors and targets.
OBJECTIVE: To clarify the effect of Jisuikang on Nogo-66 receptor NgR expression after spinal cord injury.
METHODS: 144 rats were randomly divided into six groups: sham surgery group, model group, prednisone group, high-, moderate- and low-dose Jisuikang groups (n=24). Animal models of spinal cord injury were established by the modified Allen’s method in the later five groups. Rats in the prednisone group were daily given 0.06 g/kg prednisone acetate by lavage, once a day. Rats in the high-, moderate- and low-dose Jisuikang groups were daily intragastrically given 12.5, 25 and 50 g/kg Jisuikang, once a day. Rats in the sham surgery and model groups were intragastrically daily given 20 mL of saline, once a day. Rats in each group were administered drugs until death.
RESULTS AND CONCLUSION: Compared with the model group, NgR protein and mRNA expression levels were significantly lower in the prednisone, moderate-and low-dose Jisuikang groups. These data suggested that Jisuikang can improve the recovery of neurological function after spinal cord injury and effectively inhibit NgR protein expression at the site of injury so as to suppress the microenvironment factors harmful to nerve regeneration and further improve the microenvironment of nerve regeneration. Subject headings: Drugs, Chinese Herbal; Spinal Cord Injuries; Axons; Tissue Engineering

Objective To study the role of P53 target gene TIGAR on proliferation ,migration and invasion of lung cancer cells . Methods siRNA was introduced to knock down TIGAR in A 549 cells .Proliferation was detected by CCK-8 .The ability of migra-tion and invasion was measured by and ,respectively .was used to evaluate the expression levels of related proteins .Results Knock-down of TIGAR reduced the proliferation rate (P< 0 .05) ,inhibited the ability of migration and invasion ,decreased expression levels of MMP-2 and MM P-9 .Conclusion TIGAR promotes proliferation ,migration and invasiveness of lung cancer cells .

Objective To explore the effect of Shenshuaining capsule on nutritional status and blood rheology in patients with chronic renal failure.Methods 150 cases of chronic renal failure were randomly selected and were randomly divided into observation group and control group, 75 cases in each group.The control group were given hemodialysis for treatment,and the observation group was treated with Shenshuaining capsule one week after dialysis.The clinical effect,nutritional status and biochemical indicators in the two groups were observed.Results The total effective rate in the observation group was 86.67%, which was higher than 72.00% in the control group,the difference was statistically significant(x2=4.920,P<0.05).After treatment,the nutrition indicators of HBG,ALB,IgG,IgA and IgM in the observation group were (135.7±16.4)g/L,(34.5±2.9)g/L,(10.8±0.7)g/L,(2.1±0.4)g/L and (1.7±0.3)g/L respectively, which were higher than those in the control group[(113.8±16.9)g/L,(28.4±2.5)g/L,(7.6±0.8)g/L,(1.3±0.3)g/L and (1.2±0.2)g/L], the differences were statistically significant(t=8.054,13.797,26.070,13.856,12.010,all P<0.01).In the observation group,the incidence rate of undernutrition was 25.33%, which was lower than 46.67% in the control group, the difference was statistically significant(x2=45.905,P<0.05).After treatment,the blood urea nitrogen and blood urea in the observation group were (349±64)μmol/L and (10.4±3.7)mmol/L respectively,which were lower than those in the control group[(462±51)mol/L and (14.2±4.5)mmol/L], the differences were statistically significant(t=11.958,5.649,all P<0.05).After treatment,the creatinine clearance rate and concentration of hemoglobin in the observation group were (25.1±6.1)mL/min and (92.1±21.1)g/L respectively, which were higher than those in the control group[(21.7±5.4)mL/min and (84.7±15.3)g/L], the differences were statistically significant(t=3.614,2.459,all P<0.01).After treatment, the high blood viscosity,low blood viscosity,plasma viscosity and hematocrit in the observation group were (3.59±0.94)m/Pas,(7.84±0.97)mPa/s,(1.31±0.15)mPa/s and (0.33±0.07) respectively, which were lower than those in the control group[(4.41±1.13)mPa/s,(8.72±1.05)mPa/s,(1.43±0.24)mPa/s and (0.42±0.05)], the differences were statistically significant(t=11.958,5.649,2.459,all P<0.01).Conclusion For patients with chronic renal failure,Shenshuaining capsule combined with hemodialysis can improve the nutritional status and blood rheology.

The bFGF plays an important role in embryonic development of tendons and ligaments and in the healing of injuried tendons and ligaments. The eukaryotic expression plasmid of rat basic fibroblast growth factor (bFGF) gene was constructed in order to further investigate the bFGF function in molecular regulatory mechanism in the repair of tendons and ligaments and to provide the foundation for the clinical application. The cDNA fragments of bFGF were cloned from the skin of rats by RT-PCR, and recombinated to the pMD18-T vector. The cDNA encoding bFGF was cloned from the pMD18-T vector by RT-PCR, digested with restriction enzyme EcoR I Pst I and bound to eukaryotic expression plasmid pIRES2-EGFP to construct eukaryotic expression plasmid pIRES2-EGFP-bFGF. The pIRES2-EGFP-bFGF was transfected into the tenocytes by lipid-mediated ransfection technique. MTT test was used to detect the biological activity of bFGF in supernatants after the transfection. The expression of type I and III collagen genes was detected by using RT-PCR. It was verified that the pIRES2-EGFP-bFGF was successfully constructed, and its transfection into tenocytes could significantly enhance the biological activity of bFGF, and increase the expression of type I and III collagen mRNA, suggesting that pIRES2-EGFP-mediated bFGF gene therapy was beneficial to the repair of tendons and ligaments.

Objective To explore the ziggy joint amomum six columns nai jun pill in patients with diabetic nephropathy urinary inhibition c and serum c-reactive protein levels.MethodsSelect 80 patients with diagnosis of diabetic nephropathy, randomly divided into 2 groups, 40 cases in each group.Craig column nai treatment, control group given on the basis of the observation group in the control group given six jun pill treatment, treatment of 12 weeks, before and after treatment were detected blood glucose (FBG), glycosylated hemoglobin (HbA1C), urinary inhibition c (CysC) and serum c-reactive protein (CRP) and 24 h urinary microalbumin urine trace albumin (24 hUAE) changes.ResultsCompared with before treatment, 24 hUAECysC, CRP and 2 group were lower (P< 0.05);Compared with control group, 24 huae CysC, CRP and the observation group were lower (P< 0.05), the clinical curative effect is higher (P< 0.05), and high safety.ConclusionZiggy joint amomum six columns nai jun pill treatment of the diabetic nephropathy is a effective way, and can significantly reduce serum CysC, CRP levels.

Objective To investigate the correlation between the D-dimer level and the acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods The changes of D-dimer levels in 156 patients (AECOPD group) before and after treatment were measured and compared with the D-dimer levels of 156 healthy subjects(control group).Results The concentration of plasma D-dimer in the AECOPD group was (1.35 ± 1.46) mg/L,which was significantly higher than (0.25 ± 0.16)mg/L in the control group,and the difference was statistically significant(t =9.354,P ＜0.05).The level of plasma D-dimer in the AECOPD group was (0.69 ± 0.87)mg/L,and compared with (1.35 ± 1.46)mg/L before treatment,the difference was statistically significant(t =4.850,P ＜ 0.05).Conclusion D-dimer level is correlated to the condition of patients with chronic obstructive pulmonary disease,it can be used as one of the indicators to diagnose whether a patient is in hypercoagnlable state or suffers thrombotic diseases.