Objective To supply large amount of seed cells for the cartilaginous tissue engineering,adult adipose-derived stem cells(ADSCs) were differentiated directly into cartilaginous cells.Methods Adipose tissue donated voluntarily by the adult patients after liposuction was isolated and subcultured.The cell activity was detected by methyl thiazolyl tetrazolium(MTT) assay and the proliferation curve line was drawn.The cultured stem cells were identified with flow cytometry.After the subculturing,the fourth generation of ADSCs were differentiated directly into cartilaginous cells,and then were marked by Alcian blue.Type II collagen expression in the ADSCs after cartilaginous differentiation was examined by immunohistochemical assay.Results ADSCs proliferated in vitro fast,and kept a stable multiplication till the 13th~15th generation.The results of flow cytometry showed that the cultured ADSCs had the specific features of the stem cells,and then were differentiated into cartilaginous cells.Positive Alcian blue staining and type II collagen expression indicated that the obtained cells had the function of normal cartilaginous cells.Conclusion ADSCs are differentiated directly into cartilaginous cells successfully,which will supply seed cells for the cartilaginous tissue engineering.

Objective:To explore the application of Tongdu Tiaoshen acupuncture in patients with hemiplegia after stroke.Methods:Randomized controlled trial. A total of 70 patients who met the inclusion criteria for post-stroke hemiplegia in our hospital from January 2020 to December 2021 were divided into two groups, with 35 cases in each group, according to the random number table. The control group was given conventional treatment combined with rehabilitation exercise therapy, and the observation group was given Tongdu Tiaoshen acupuncture on the basis of the above, and both groups were treated continuously for 1 month. Compare of the two groups Fugl-Meyer Assessment (FMA), Modified Ashworth Scale (MAS), National Institutes of Health Stroke Scale (NIHSS), Activity of Daily Living (ADL) score; Brunnstrom stage method was used to assess the patient's motor ability, and Holden walking function grade was used to assess the patient's walking ability. The activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time(TT), and serum fibrinogen (FIB), D-dimer (D-D), plasminogen activator inhibitor 1 (PAI-1) levels were observed by ELISA. Adverse events were recorded and clinical efficacy was evaluated.Results:The total effective rates of the observation group and the control group were 97.14% (34/35) and 77.14% (27/35), respectively, and the difference between the two groups was statistically significant ( χ2=6.25, P=0.012). After treatment, the scores of FMA and ADL in the observation group were significantly higher than those in the control group ( t values were 9.23 and 9.54, respectively, P<0.01), the MAS and NIHSS scores were significantly lower than those in the control group ( t values were 10.23 and 11.97, respectively, P<0.01). After treatment, the distribution of patients in Brunnstrom stage Ⅴ and Ⅵand Holden functional walking stage Ⅳ and Ⅴ in the observation group was significantly higher than those in the control group ( χ2 values were 11.96 and 11.27, respectively, P<0.05). After treatment, APTT, PT and TT in the observation group were significantly longer than those in the control group ( t values were 10.37, 13.57 and 6.54, respectively, P<0.01); serum FIB, D-D and PAI-1 levels were significantly lower than those in the control group ( t values were 12.85, 11.94 and 27.39, respectively, P<0.01). No adverse reactions occurred in both groups. Conclusion:The treatment of post-stroke hemiplegia with Tongdu Tiaoshen acupuncture can improve the effect of clinical treatment and the ability of daily life, neurological function, limb function, movement and walking ability, and stabilize the clotting state.