Objective To observe the therapeutic effect of plum-blossom needle therapy and cupping therapy combined with traction on cervical spondylosis of vertebral artery type.Methods 60 patients with cervical spondylosis of vertebral artery type were randomly divided into the treatment group and control group with 30 cases in each group. The patients of the treatment group were treated with plum-blossom needle therapy and cupping therapy combined with traction, and those of the control group only with traction. The therapeutic effects of two groups were compared.Results The total effective rate of the treatment group was 83.3 %, that of the control group was 60.0%. There was a significant difference between two groups ( P<0.05).Conclusion The effect of plum-blossom needle therapy and cupping therapy combined with traction on cervical spondylosis of vertebral artery type is superior to traction therapy alone.

To optimize the growth condition for the established gene engineer bacteria express cardiac troponin-I(cTnI) and to obtain purified cTnI as an antigen to produce clinical assay kits used in acute myocardial injury(AMI) diagnosis.Plackett Burman Design(PBD) was applied to select the factors which effect the expression of cTnI in Escherichial coli(E.coli) mostly.Induction time,pH and KCl were proved influenced expression of cTnI notably.Afterward,Response Surface Methodology(RSM) as second step to optimize the selected three factors,an equation was deduced to predict the percent of cTnI.In the most optimized condition,the percent of cTnI can reach to 26% of total cell protein.The procedures of purification included ammonium sulfate deposition and DEAE Cellulose ion exchange chromatography.SDS-PAGE shows that purified cTnI contain one band.cTnI could be used to immune animals as an antigen to produce monoclonal antibodies with high affinity and specificity.It maybe as calibrators to harmony the difference assays of cTnI measurement in clinical.

ObjectiveTo investigate Schwann cells whether survival and migration after transplanted to central nervous system for a long-term.MethodsThe Schwann cells of rat were expended in vitro, the part of them were labeled with 5'-bromodeoxyuridine (BrdU) and transplanted to rat's middle brain injured by electric needle stimulus, the others were labeled with Hoechst 33342, seeded to PLGA scaffold, and transplanted to rat's transected spinal cord. 8 and 11 months later, rat brain and spinal cord were taken out respectively, examined by BrdU immunohistochemistry and fluorescence microscope.ResultsBrdU positive cells could be seen after 8 months and migrated toward cerebral cortex. Hoechst 33342 positive cells could be identified in scaffold and transected spinal cord after 11 months under fluorescence microscope.ConclusionGrafted Schwann cells can survive in central nervous system for a long-term and migrate toward distance.

In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.

The effects of different physical and chemical factors on hairy root growth and flavonoids production were studied in suspension culture of Saussurea medusa hairy root in 1/2 MS medium. The results showed that the following culture conditions, nitrogen concentratiaon (involved NH4+ and NO3-), 30 mmol/L; the ratio of ammonium to nitrate, 5:25; the combination of 2% sucrose and 3% glucose; 0.5 mg/L GA3; 0.5 mg/L IBA; initial pH 5.8; light cycle, 18 h/d (3500lx); temperature, 24 degrees C; shaker revolutions per minute, 100 r/min, were favourable to hairy root growth and flavonoids production. Under the above culture conditions, up to 12.8 g/L (DW) of hairy root and 1922 mg/L of flavonoids were obtained after 21 days of culture. The content of total flavonoids in hairy root was 15%, which was about 25 times as that in the wild plantlet.
Culture Media ; Flavonoids ; biosynthesis ; Plant Roots ; growth & development ; metabolism ; Saussurea ; growth & development ; metabolism ; Temperature ; Tissue Culture Techniques

Objective To explore the expression of hypoxia-inducible factor-1?(HIF-1?) protein and its mRNA in cultured cortical neurons after hypoxia,ischemia or hypoxia-ischemia(HI) and explore the possibilities of HIF-1? gene therapy in HI neurons.Methods The in vitro models of HI,pure hypoxia and pure ischemia were established using embryonic day 16-18 rats cortical neurons.Immunohistochemical and in-situ hybridization were performed to examine the expression of HIF-1? protein and its mRNA at different reperfusion time points in neurons.Results The expression of HIF-1? protein was very week in normoxic cultured neurons,but was up-regulated while treated with hypoxia and(or) ischemia.HIF-1? expression reached peak at 4 to 8 h after reperfusion with HI,which were statistically significant higher than that at other time points(Pa=0),and decreased gradually at 12 h.Furthermore,HIF-1? protein expression was significantly higher in HI group compared with that in the pure hypoxia or ischemia group(Pa=0).HIF-1? mRNA reached peak immediately after HI,decreased gradually at 2 h,and returned to the baseline at 8 h after reperfusion.Conclusions HIF-1? expression on cortical neurons is regulated differently with hypoxia,ischemia or HI treatment,HIF-1? gene therapy for HI neurons maybe a useful method in the future studies.

OBJECTIVETo observe the differences of the expression of Cx43 in rats' acupoints and sham-acupoint when needled blocks, and explore the relatively specificity of acupoints.

METHODTwenty Wistar rats were randomly divided into two groups: an acupuncture group and a control group, ten in each group. The rats in the control group were not treated and sacrificed directly to by air embolism; the rats in the acupuncture group were needled at "Neiguan (PC 6)", "Zusanli (ST 36)", "Dazhui (GV 14)" and "Guanyuan (CV 4)", Sham-acupoints were on 1 cm next to each of the above acupoints and "Yongquan (KI 1)" were needled by hand manipulation, retented needles for 30 min, then all the rats were sacrificed by air embolism. With the rats fixed, the tissue blocks at the acupoints and sham-acupoints were collected, then quick-frozen and cut into sections. The sections were stained by immunohistochemical method (Cx43).

RESULTSThe expression of the rats' Cx43 at the acupoints was significantly higher than that at the non-points in both groups (all P < 0.01). The expression of Cx43 at the acupoints in the acupuncture group was significantly higher than that in the control group (all P < 0.01).

CONCLUSIONThe expression of Cx43 at acupoints and non-points is significantly different, which indicates that the acupoints are actually existed and have relatively specificity.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Connexin 43 ; genetics ; metabolism ; Gene Expression Regulation ; Humans ; Male ; Rats ; Rats, Wistar

OBJECTIVE: To bring forward a new method for automatic measurement of the translation and rotation of the eye movement. METHODS: A centroid method, edge filter, and ellipse fitting were used to get the accurate position of the eye center. The rotation angle of the eye was determined by Fourier-Translation arithmetic operators. RESULTS: The simulation image test and initial clinical experiment obtained a good precision. CONCLUSION This method can eliminate the influence of eyelid overlapping and illumination, which can measure the 3-dimensional eye movement accurately.
Electronic Data Processing ; Electronystagmography ; methods ; Eye Movements ; Humans ; Imaging, Three-Dimensional ; methods ; Models, Anatomic ; Models, Biological ; Nystagmus, Optokinetic ; physiology ; Reflex, Vestibulo-Ocular ; physiology ; Video Recording ; Vision, Ocular

Objective:To establish a human lung squamous carcinoma cell line and to study its biological characteristics. Methods:Lung squamous carcinoma specimens were freshly resected during operation;the tissues were incubated in vitro and the cell line was named CHLH-1.The biological characteristics of the cells were studied by light microscopy,electron microsco- py,chromosome analysis and transplantation experiment.Results:Cells from the specimens of the primary tumor,the CHLC- 1 cell line and the cells from transplanted tumor possessed the characteristics of malignant squamous epithelium under light and electron microscope.The cell growth curve,doubling time and mitotic index were also observed in vitro.Nuclear chromosome analysis revealed that the tumor was a subtriploid with a mode of 60-68 per cell.Tumor nodes were observed under the skin of nude mice by heterogenic transplantation.Conclusion:The characteristics of the established cell line suggest that it is a newly established human squamous carcinoma cell line.

OBJECTIVETo investigate the biological characters of human skin fibroblasts in fibroblast populated collagen lattice (FPCL).

METHODSThe human fibroblasts were cultured in 3D and the collagen of the rat tail was also prepared. They were examined with the comprising cell cycle and apoptosis, mRNA expression of TGF beta1, and fibronectin, and cell morphology.

RESULTSThe flow cytometry showed that the G0/G1, stage cells were 79% +/- 3%, 87% +/- 2% after the 7 days and 14 days separately, and there were not apoptosis peak observed. RT-PCR analysis revealed that the mRNA expression of TGF beta1, and fibronectin had no difference between human skin fibroblasts cultured in 3D and 2D. Electron microscope showed the cells were plenty of chromatin and organelles.

CONCLUSIONSThe proliferation of the human skin fibroblasts in FPCL is slow, but its biological viability is better.

Animals ; Cell Culture Techniques ; Cell Division ; Cells, Cultured ; Collagen ; Extracellular Matrix ; Fibroblasts ; cytology ; Humans ; Rats ; Skin ; cytology ; Tissue Engineering ; methods