Objective To explore the effects of calcitonin gene-related peptide (CGRP) on proliferation of vascular endothelial cells (VECs) and osteogenic differentiation of bone mesenchymal stem cells (BMSCs) of osteoporotic rats in vitro,and cell/ molecular mechanism of promoting osteoporotic fracture healing.Methods 10 3-month-old SD rats were employed to make osteoporosis model by taking bilateral ovariectomy.Micro-CT analysis was used to determine the success of modeling at 6 months of postoperation.BMSCs and VECs were isolated from osteoporotic rats by blood adherence mcthod.10-7-10-10 mol/L CGRP were added to the culture medium.CCK-8 method was used to examine the cell proliferation.Effect of CGRP on osteogenic differentiation of BMSCs:Alkaline phosphatase (ALP) activity,and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of ALP,Collagen Ⅰ,BMP-2,Osteonectin and RunX2 gene.Effect of CGRP on vascularization of VECs:Vascular cell endothelial growth factor (VEGF) protein secretion and gene expression level were tested by ELISA and RT-PCR.Resuits BMSCs and VECs of osteoporotic rats were isolated successfully.The optical density (OD) values of CCK-8 method increased in a dose-dependent manner,for BMSCs at 7 d,the control group of OD value was 0.58±0.02,the CGRP group 10-7-10-10 mo]/L was to 0.80±0.03,0.79±0.03,0.74±0.03,0.69±0.03,difference was significant;for VECs,the control group of OD value was 2.20±0.01,the CGRP group 10-7-10-10 mol/L was to 4.12±0.04,3.90±0.02,3.56±0.04,3.12±0.04,difference was significant.Effect of CGRP on osteogenic differentiation of BMSCs:Alkaline phosphatase (ALP) activity,and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of ALP,Collagen Ⅰ,BMP-2,Osteonectin and RunX2 gene.Effect of CGRP on vascularization of VECs:Vascular cell endothelial growth factor (VEGF) protein secretion and gene expression level were tested by ELISA and RT-PCR,difference between the control group and CGRP group were all significant.Conclusion 10-7 mol/L,10-10 mol/L of CGRP can directly promote the proliferation of the BMSCs and VECs of osteoporotic rats,promote the osteogenic differentiation of BMSCs,improve the expression of VEGF gene and protein secretion,promote angiogenesis.

Objective To investigate the effect of improving immunity in the treatment of chronic brucellosis,and to analyze and evaluate its clinical curative effect.Methods A patient with chronic brucellosis was treated with Mongolian medicine combined with chemical drugs to enhance immunity.The clinical symptoms,serological antibodies,Brucella DNA and immune function were compared before and after treatment.The specific antibody against Brucella in serum was detected by tube agglutination test (SAT) and tiger red plate agglutination test (RBPT).Brucella DNA in serum and blood cells was detected by PCR,and the peripheral blood lymphocyte subsets were detected by flow cytometry.Immuno-luminescence technique was used to detect serum immunoglobulin and complement components.Results After treatment,the clinical symptoms such as cold back,fatigue,and joint pain disappeared completely,and the results of serum specific antibodies against Brucella were SAT 1 ∶ 50 (++)and RBPT (+) with no changes before and after treatment,and the results of cells and serum were both negative after treatment though the results of DNA detection of Brucella were cell positive and seronegative before treatment.The results of immunological function test showed that γδT cells decreased to 9.50％ after treatment compared to 14.00％ before treatment,and the percentage of monocytes and Treg cells were 5.59％ and 7.33％ after treatment,which were higher than 3.35％ and 4.72％ of before treatment,and the level of complement C3 was 0.79 g/L before treatment and 0.91 g/L after treatment that was returned to normal reference range (0.88 ～ 2.01 g/L).Conclusion The patients with chronic brucellosis can improve their clinical treatment by improving immunity.

Objective: To understand the association between non suicidal self injury and self control among rural junior high school students in Guizhou Province, and to provide reference for the development of prevention and control measures. Methods: Using a multistage stratified random cluster sampling method, a questionnaire survey was administered to 4 452 junior high school students in 30 rural schools in Guizhou Province from November to December 2021 to investigate non suicidal self injury and self control using the Non suicidal Self Injury Scale and the Self Control Scale. Results: A total of 986 (22.2%) students had occasional non suicidal self injurious behaviors, 593 (13.3%) students reported frequent non suicidal self injury, and the occurrence of non suicidal self injury among junior high school students varied significantly by gender, grade, only child, residence, left behind experiences, academic performance, academic pressure, parent child relationship, peer relationship, teacher student relationship, and exposure to domestic violence ( χ 2=6.97-168.68, P <0.05); the average score of self control among junior high school students in rural Guizhou Province was 48.0 (39.0, 57.0), and the differences in self control scores, impulse control, healthy habits, resist temptation, focus on work, and moderate entertainment were of statistical significance among students with varied frequency of self injurious behavior ( Z=612.08, 464.64, 193.10, 228.86, 194.04 , 542.20, P <0.05); high self control score was associated with more non suicidal self injurious behavior ( OR=1.06, 95%CI=1.05-1.07, P <0.05). Conclusion The prevalence of non suicidal self injury is high among rural junior high school students in Guizhou province, and self control shows a predictive effect on non suicidal self injurious behavior.Schools, families and relevant government departments should pay full attention to it and formulate targeted prevention and control measures.

Objective We aimed to compared matrix metalloproteinase-13 (MMP-13) and transforming growth factor-β1 (TGF-β1) in synovial fluid,the phosphorylation level of Smad3 in articular cartilage (P-Smad3),and their correlation with traditional Chinese medicine (TCM) patterns in patients with knee osteoarthritis (KOA) of liver-kidney deficiency pattern and pattern of intermingled phlegm and blood stasis.Methods Using a cross-sectional field investigation method,KOA patients hospitalized in the Orthopedics Department of Dongzhimen Hospital,Beijing University of Chinese Medicine from September 2019 to February 2023 were collected. A total of 112 KOA patients were included,among which 63 cases were diagnosed with liver-kidney deficiency pattern,and 49 cases were diagnosed with pattern of intermingled phlegm and blood stasis. The intensity of knee pain,function,and X-ray imaging result were quantified using the Visual Analogue Scale (VAS),Lysholm Knee Scoring Scale,and Kellgren-Lawrence (K-L) Grading Scale,respectively. The TCM pattern was identified and quantified using a TCM Pattern Scoring Scale. Immunohistochemistry was used to determine the phosphorylation characteristics of Smad3 in articular cartilage,and ELISA was used to measure the contents of MMP-13 and TGF-β1 in synovial fluid. The level characteristics and their correlation with the degree of syndrome were analyzed.Results (i) There was no statistically significant difference in VAS scores,Lysholm scores,and K-L grades between KOA patients with different TCM patterns. (ii) Compared with KOA patients with pattern of intermingled phlegm and blood stasis,patients with pattern of liver-kidney deficiency had higher levels of MMP-13 in synovial fluid and lower levels of TGF-β1 in synovial fluid (P<0.05). (iii) In KOA patients with liver-kidney deficiency pattern,there was a positive correlation between the level of MMP-13 in synovial fluid and the score of TCM pattern (r=0.292,P=0.020),while there was a negative correlation between the level of TGF-β1 in synovial fluid and the score of TCM pattern (r=-0.781,P<0.001). In KOA patients with pattern of intermingled phlegm and blood stasis,there was also a positive correlation between the level of MMP-13 in synovial fluid and the score of TCM pattern (r=0.936,P<0.001). (iv) The mean optical density value of P-Smad3 in articular cartilage was lower in KOA patients with liver-kidney deficiency pattern than in pattern of intermingled phlegm and blood stasis (P<0.05).Conclusion KOA patients with liver-kidney deficiency pattern or pattern of intermingled phlegm and blood stasis have different levels of TGF-β1 and MMP-13 in synovial fluid,as well as varying degrees of Smad3 phosphorylation in articular cartilage,which is consistent with the analysis of etiology and pathogenesis under different patterns. The levels of TGF-β1 and MMP-13 in synovial fluid of patients with liver-kidney deficiency pattern can reflect the severity of the pattern to a certain extent,and the mechanism may be related to the inhibition of the activation level of the TGF-β/Smad signaling pathway. This study enriches the research content of the material basis of TCM patterns.