Objective To obtain the Tat-GFP fusion proteins with penetrating activity and labeled with green fluorescence protein (GFP), and to explore the cell membrane penetrating activity of Tat-GFP in MCF-7 cells. Methods The plasmid pET-24a-Tat-GFP was transformed into Escherichia coli BL21 cells. Different concentrations (0.5 and 1.0 mmol · L-1 ) of isopropyl-β-D-thiogalactopyranoside (IPTG ) and cell culture temperatures (22℃ and 37℃)were used to optimize the protein expression.The Tat-GFP proteins in supernatant were purified using Ni-IDA resins. Western blotting analysis was used to identify the Tat-GFP protein, and confocal laser scanning microscope (CLSM ) was used to examine the cell penetration of Tat-GFP protein. Results There was no significant difference in the Tat-GFP protein production induced by 0.5 and 1.0 mmol·L-1 IPTG;however,the low temperature (22℃)-induced BL21 cells expressed more Tat-GFP proteins than that at 37℃ induction.The Western blotting analysis results showed that GFP antibody could specifically recognize the proteins in PVDF membranes in dose-dependent manner;the CLSM results indicated the distribution of green fluorescence in cytoplasm and nucleus of MCF-7 cells.Conclusion The Tat-GFP protein highly expresses in the supenatant of Escherichia coli i BL2 1 cells at low temperature;the obtained Tat-GFP protein with green fluorescence preserves the cell penetrating activity.

Objective To investigate the clinical efficacy of a new-type moxibustion box in treating cervical spondylotic radiculopathy.Methods Sixty patients with cervical spondylotic radiculopathy were randomly allocated to treatment and control groups, 30 cases each. The treatment group received nape-eight-needle acupuncture plus new-type moxibustion box therapy and the control group, nape-eight-needle acupuncture plus conventional moxibustion box therapy. After 10 treatments, the NPQ score, the symptom and sign score and the VAS score were recorded in the two groups and the clinical therapeutic effects were compared between the two groups.Results The total effective rate was 100.0% in the treatment group versus 83.3% in the control group, and the difference was statistically significant (P<0.05). There were statistically significant pre-/post-treatment differences in the NPQ score, the symptom and sign score and the VAS score in the two groups (P<0.01,P<0.05). There were statistically significant post-treatment differences in the NPQ score, the symptom and sign score and the VAS score between the treatment and control groups (P<0.05).ConclusionNape-eight-needle acupuncture plus a new-type moxibustion box is an effective way to treat cervical spondylotic radiculopathy.

OBJECTIVE: To investigate the stability of 0.5% glutaraldehyde solution. METHODS: Classic constant temperature accelerated test method and sample observation method were applied with the content of glutaraldehyde as index. The content of glutaraldehyde was determined by titration which was determined by titration. RESULTS: The content change of 0.5% glutaraldehyde solution was in line with the first order kinetics, and the results of both methods were similar. The validity duration of 0.5% glutaraldehyde solution was 85.7 days at room temperature (25 ℃). CONCLUSION: 0.5% glutaraldehyde solution should be used up as soon as possible and the perfect time is less than 2 week.

Objective To design an exoskeleton mechanical device for hand dysfunction rehabilitation. Methods Based on the bionic　principles of the motion and the structure of finger joints, a micro exoskeleton mechanical structure with multi-joint fingers and multi-DOF　motion was established, which included wearable finger exoskeleton and micro-motor drive mechanism. With the motion analysis software,the motion simulation analysis of the design was carried out on the basis of the three-dimensional model of the mechanical device. Results　and Conclusion The structure of training device is compact and light, and the exoskeleton finger mechanism can move normally in the　range of 0～67° without interference. Simulation data show that the mechanical device can complete planning exercises, and meet the design　requirements of kinematics and mechanical properties.

Animals ; Bone and Bones ; cytology ; Cell Differentiation ; Mesenchymal Stromal Cells ; drug effects ; physiology ; Neurons ; physiology ; Plant Extracts ; pharmacology ; Rats ; Salvia miltiorrhiza ; chemistry

In most mammalian central nervous system diseases, axons are damaged.Due to the limited ability of damaged neurons to promote axonal regeneration, the formation of glial scar and the release of inhibitory nutrients, it is difficult to regenerate axons of damaged neurons. The purpose of this study was to investigate the effect of cerebroprotein hydrolysate for injection (II) (CBL) on neuritogenesis and its underlying mechanism. Immunofluorescence staining was used to detect the axon length of mouse neuroma cells (Neuro-2a) and mouse primary cortical neuronal cells. Western blotting was used to detect the expression of phosphorylated TrkB protein in Neuro-2a cells and mouse primary cortical neuronal cells. The results showed that CBL could increase the axon length of Neuro-2a cells or mouse primary cortical neuronal cells, and that the phosphorylation level of TrkB in neuronal cells was significantly increased when 5 μg/mL CBL was applied to neuronal cells for 1 h. In conclusion, CBL can promote neuritogenesis, and increase the expression of phosphorylated TrkB, which may be related to the activation of TrkB signaling pathway.

Objective To research the influenza virus infection on rat vascular smooth cells number,proliferation,apoptosis,the amount of IL-6,sFas,platelet-derived growth factor(PDGF) and the mechanism of atherosclerosis.Methods Flow cytometry,enzyme-linked immunosorbent assay and cell count experiments were used to detect these indicators at 0 h,6 h,12 h,24 h,48 h.Results After influenza virus infected at 0 h,proliferation,apoptosis condition were 10.39％,0.44％,respectively; at 6 h,proliferation,apoptosis respectively increased to 12.68％,0.73％ ; proliferation reached the peak at 12 h (18.01％),instead apoptosis decreased to 0.14％ ; at 24 h,proliferation decreased to 12.89％ and apoptosis markedly increased to 1.09％ ; at 48 h,proliferation further reduced to 7.07％ and apoptosis reached the peak(4.61％).The number of cells and the cytokine secretion were statistically significant to control group (P＜0.05).Conclusion Influenza virus infection might lead to change of cell proliferation and apoptosis and involve the atherosclerosis form and development,and cytokines played an important role in them.

Objective To construct single chain antibody specific to membrane protein of Schistosoma japonicum by gonetic engineering technique. Methods The V\-H (heavy-chain variable region) and V\-L (light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using V\-H and V\-L genes, then expressed by IPTG. Results The V\-H and V\-L genes were obtained through PCR. The DNA sequences showed that V\-H and V\-L were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin-fused proteins about 36\^2 kDa. Conclusion A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed.

The protoplasts of Micromonospora purpurea,the high yield strains of gentammicin producer were mutagenized by diethyl sulfate(DES)and ultraviolet radiation(UV)respectively,then fused,screened by gentamicin resistance and regenerated.The average fermentation unit 2200?U/ml could be achieved by shake flask for 10 batches.The average fermentation unit 1900?U/ml could be obtained by 5L fermentor for 7 batches.The quality of the end product conformed to CP2000,BP2000 and USP26 pharmacopoeia.

Objective To observe the changes of abdominal oxygen saturation in very low birth weight infants (VLBWI)with feeding intolerance (FI)within 1 4 days after birth monitored by near infrared spectroscopy (NIRS).Methods VLBWI fitting entry criteria were enrolled into this study.NIRS monitoring was carried out to detect cerebral oxygen saturation (ScO2 )and abdominal oxygen saturation (SsO2 ).Data were analyzed between FI infants and feeding tolerance (FT)infants.FI was defined as follows:gastric residual of more than 50% of the previous feeding volume;emesis or abdominal distention or both;decrease,delay or discontinuation of enteral feedings. Results 93 VLBWI were enrolled.52 cases(55.91 %)presented with FI,including 29 cases(31 .1 9%)of gastric residual increasing and 23 cases(24.73%)of emesis with or without abdominal distention within 1 4 days after birth. The levels of SsO2 and SsO2 /ScO2 showed no differences in infants with FT and with FI within 24h after birth (P >0.05).The change rates of the median of SsO2 and SsO2 /ScO2 in FT infants were similar during 1 4 days (P >0.05).While both the change rates of SsO2 and SsO2 /ScO2 were markedly decreased 1 day before and the day of FI (P <0.01 ).The decreasing degree of SsO2 was similar between infants with gastric residual increasing and infants with emesis with or without abdominal distention[(1 6.2 ±5.1 )vs (1 7.4 ±3.6)%,t =0.733,P =0.476]. Conclusion Abdominal oxygen saturation measured by NIRS may be a useful method for infants adjusting the feeding plan.