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Objective @#To explore and optimize the in vitro primary culture method of astrocytes in neonatal mouse cerebral cortex , which provides a better solution for the in vitro culture of astrocytes.@*Methods @#In order to optimize the in vitro culture method of mouse cerebral cortex astrocytes , 3 ⁃day⁃old C57BL/6J mouse cerebral cortex tissues were taken , meninges and blood vessels were removed , digested by pancreatic enzymes and centrifuged , andhigh⁃glucose dulbecco ′s modified eagle medium (DMEM) was added to form cell suspension , which was purified by differential adhesion method , cross hand method and constant temperature shaking method.The cells were inoculated in poly⁃D ⁃lysine⁃coated culture bottles with different culture densities , and the purity of astrocytes was determined by morphological ob servation and immunofluorescence staining.@*Results @#The cells were inoculated at a density of 5 × 106 cells per bottle with good effect and high activity. The purity of astrocytes reached 99% by using high sugar DMEM medium combined with differential adhesion method , cross hand method and constant temperature shaking method.@*Conclusion @#The primary culture method of astrocytes in mouse cerebral cortex is successfully established and optimized.
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Objective@#To explore and optimize the primary culture method of neonatal mouse hippocampal neurons in vitro．To construct a G-protein-coupled receptor kinase 2 ( GRK2) knockout HT22 cell line．@*Methods @#Hippocampal tissue of C57BL6 /J mice on day 1－2 was taken，digested with trypsin and pipetted to form a cell suspension，and supplement was added to Neurobasal-A medium to maintain cell growth． CRSIPR / Cas9 gene editing technique was used to construct HT22-GRK2 －/ － cell line，and the knockout efficiency of GRK2 was detected by immunofluorescence staining and Western blot. @*Results @#Primary hippocampal neurons of newborn mice were put into six-well plates with 3 × 107 /well using a serum-free culture method，which could get a high purity and good activity ; HT22-GRK2 －/ － cell line was constructed successfully．@*Conclusion@#The primary culture method of mouse hippocampal neurons was successfully established and optimized，and HT22-GRK2 －/ － cell line was successfully constructed by CRSIPR / Cas9 gene editing technique.
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Objective To analyze the therapeutic effect ofShengji-Yuhong ointment in the treatment of patients with pressure ulcer.Methods 86 patients of pressure ulcer were randomly divided into a control group and an observation group, with 43 cases in each. After debridement, the wound was covered with vaseline gauze in the control group, whileShengji-Yuhong ointment in the treatment group. 10 days constituted 1 course of treatment, and both groups were treated for 3 courses. The blood supply of the whole blood viscosity, plasma viscosity, erythrocyte aggregation index detection; white blood cell count (WBC), erythrocyte sedimentation rate (ESR), and C reactive protein (CRP) were observed in order to observe the control condition of the patients with wound infection.Results The total effective rate was 95.3% (41/43) and 74.4% (32/43) in the observation group and control group respectively, with significant difference between two groups (χ2=5.800,P=0.016). After treatment, the whole blood viscosity (high-shea) (4.06 ± 1.38 mPa?svs. 4.74 ± 1.62 mPa?s,t=2.095), the whole blood viscosity (medium-shea) (3.71 ± 1.22 mPa?svs. 4.34 ± 1.41 mPa?s,t =2.216), blood reduction viscosity (1.13 ± 0.22 mPa?svs.1.44 ± 0.51 mPa?s,t=3.660), the whole blood viscosity (medium-shea) (4.16 ± 0.48 mPa?svs. 4.51 ± 0.89 mPa?s,t=2.270) obviously compared with group before treatment decreased (P<0.05). The patients in the observation group in the whole blood viscosity (medium-shea) (3.71 ± 1.22 mPa?svs. 4.16 ± 0.48 mPa?s,t=2.251), and blood reduction viscosity (1.13 ± 0.22 mPa?svs. 1.32 + 0.31 mPa?s,t=3.278) in the observation group were obvious better than the control group (P<0.05). After the treatment the WBC, CRP, ESR in the observation group were decreased significantly than the control group (t=5.947, 7.198, 12.064,P<0.01).ConclusionShengji-Yuhong ointment can effectively control the PU infection in the wound, improve wound tissue under the blood circulation, and promote wound healing.
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ObjectiveTo evaluate the curative effect of local massage with safflower oil in patients with phlebitis following peripheral intravenous catheters.MethodsA total of 71 patients with phlebitis following peripheral intravenous catheters wererandomly divided into 2 groups, a safflower oil group with 36 cases, and a magnesium sulfate group with 35 cases. The magnesium sulfate group was treated by local external application of 33% magnesium sulfate on phlebitis at the puncture site, while the safflower oil group was treated by external application of safflower oil 3~5 cm around the peripheral vein puncture site and massage. Both groups were treated for 48 h. Visual Analog Scale (VAS) was used to assess pain degree, marking method to label the localred swelling area.ResultsThe VAS score (0.81±0.13vs.0.94±0.11;t=4.543,P<0.01) at 48 h after the treatment, and the local red swelling area at 24 h (3.62±1.22 cm2vs.4.42±1.72 cm2;t=2.335, P=0.022) and 48 h (1.07±0.25 cm2vs.3.26±1.07cm2;t=11.952,P<0.01) after the treatment in the safflower oil group were significant lower or smaller than the magnesium sulfate group.ConclusionsLocal massage with safflower oil can effectively alleviate the severity of phlebitis, relieve symptoms, reduce the score of VAS and local red swelling area, and promote the damaged tissue to restore normal.