BACKGROUND:Bone marrow mesenchymal stem cells play a crucial role in treatment of diseases,such as femoral head necrosis,and the therapeutic efficacy is closely related to the quality of the cells.The empowerment of cells has become a research focus.OBJECTIVE:To investigate the effects of hypoxia mimetic dimethylglyoxal glycine pretreatment on mitophagy and differentiation capacity of human bone marrow mesenchymal stem cells.METHODS:Bone marrow mesenchymal stem cells were extracted from the bone marrow of patients' iliac crest and cultured in vitro to the third passage.The cells were treated with dimethylglyoxal glycine at 0,10,50,and 100 μmol/Lfor 24 hours,followed by the replacement with an osteogenic induction differentiation medium,which constituted the pretreatment group.The continuous treatment group was cultured directly in osteogenic induction medium containing 0,10,50,and 100 μmol/L dimethylglyoxal glycine after cell adhesion.After 7 days of induction,alkaline phosphatase staining was performed to select the most favorable conditions for osteogenic differentiation for subsequent experiments,with normally cultured bone marrow mesenchymal stem cells serving as the control group.Alkaline phosphatase staining,alkaline phosphatase activity,oil red O staining,and related RT-qPCR were used to evaluate the osteogenic and adipogenic differentiation differences of bone marrow mesenchymal stem cells between the two groups.MitoSox staining was used to detect mitochondrial reactive oxygen species levels.Mito-tracker and Lyso-tracker staining were used to detect the co-localization of mitochondria and lysosomes.The fluorescent probe JC-1 was used to measure mitochondrial membrane potential.RESULTS AND CONCLUSION:Alkaline phosphatase staining indicated that the most beneficial treatment for bone marrow mesenchymal stem cell osteogenesis was pretreatment with 10 μmol/L dimethylglyoxal glycine for 24 hours.Compared with the control group,the experimental group showed enhanced alkaline phosphatase staining expression,increased alkaline phosphatase activity and osteogenic gene expression,reduced lipid droplet formation and adipogenic gene expression as indicated by oil red O staining,decreased mitochondrial reactive oxygen species production,increased co-localization of mitochondria and lysosomes,and elevated mitochondrial membrane potential.The results suggest that 10 μmol/L dimethylglyoxal glycine pretreatment can promote osteogenic differentiation of bone marrow mesenchymal stem cells,inhibit adipogenic differentiation,and enhance mitophagy.

BACKGROUND:Bone marrow mesenchymal stem cells play a crucial role in treatment of diseases,such as femoral head necrosis,and the therapeutic efficacy is closely related to the quality of the cells.The empowerment of cells has become a research focus.OBJECTIVE:To investigate the effects of hypoxia mimetic dimethylglyoxal glycine pretreatment on mitophagy and differentiation capacity of human bone marrow mesenchymal stem cells.METHODS:Bone marrow mesenchymal stem cells were extracted from the bone marrow of patients' iliac crest and cultured in vitro to the third passage.The cells were treated with dimethylglyoxal glycine at 0,10,50,and 100 μmol/Lfor 24 hours,followed by the replacement with an osteogenic induction differentiation medium,which constituted the pretreatment group.The continuous treatment group was cultured directly in osteogenic induction medium containing 0,10,50,and 100 μmol/L dimethylglyoxal glycine after cell adhesion.After 7 days of induction,alkaline phosphatase staining was performed to select the most favorable conditions for osteogenic differentiation for subsequent experiments,with normally cultured bone marrow mesenchymal stem cells serving as the control group.Alkaline phosphatase staining,alkaline phosphatase activity,oil red O staining,and related RT-qPCR were used to evaluate the osteogenic and adipogenic differentiation differences of bone marrow mesenchymal stem cells between the two groups.MitoSox staining was used to detect mitochondrial reactive oxygen species levels.Mito-tracker and Lyso-tracker staining were used to detect the co-localization of mitochondria and lysosomes.The fluorescent probe JC-1 was used to measure mitochondrial membrane potential.RESULTS AND CONCLUSION:Alkaline phosphatase staining indicated that the most beneficial treatment for bone marrow mesenchymal stem cell osteogenesis was pretreatment with 10 μmol/L dimethylglyoxal glycine for 24 hours.Compared with the control group,the experimental group showed enhanced alkaline phosphatase staining expression,increased alkaline phosphatase activity and osteogenic gene expression,reduced lipid droplet formation and adipogenic gene expression as indicated by oil red O staining,decreased mitochondrial reactive oxygen species production,increased co-localization of mitochondria and lysosomes,and elevated mitochondrial membrane potential.The results suggest that 10 μmol/L dimethylglyoxal glycine pretreatment can promote osteogenic differentiation of bone marrow mesenchymal stem cells,inhibit adipogenic differentiation,and enhance mitophagy.

Objective: To evaluate the long term efficacy of treating the primary dysmenorrhea of cold-damp stagnation type by Acupuncture of Liji therapy. Methods: In this study, a total of 76 cases of primary dysmenorrhea of cold-damp coagulation type were randomly divided into the acupuncture of Liji therapy group, and the body acupuncture group, with38 cases in each group. Both groups were continuously treated 3 menstrual cycles, and followed up in the third and six menstrual cycles after the end of the treatment. Visual analogue scale for abdominal pain and Dysmenorrhea Symptoms scale were used as therapeutic indexes. Remove shedding cases, the long-term effects and scores of the 2 groups were compared. Besides, untoward and side effects needed to be recorded. Results: There were 2 cases lost in the acupuncture ofLiji therapy group and 1 cases in the body acupuncture group. After treatment, The clinical comprehensive efficacy of acupuncture ofLiji therapy group was better than that of body acupuncture group (P ＜ 0. 05) . Both groups of VAS scores and dysmenorrhea symptom scores were decreased to different degrees during treatment and follow-up period (P ＜ 0.05) .The follow-up data of the body acupuncture group after six menstrual cycles were higher than that of the third menstrual cycles after treatment. Compared with the two groups, the acupuncture of Liji therapy group was superior to the body acupuncture group during the third menstrual cycles follow-up (P ＜ 0.05) and six menstrual cycles follow-up (P ＜ 0.01) .The treatment satisfaction of acupuncture of Liji therapy and body acupuncture was 91.67％ and 72.97％. There were no adverse reactions in the two groups during the study period. Conclusion: Acupuncture of Liji therapy can effectively relieve dysmenorrhea symptoms and the general discomfort caused by dysmenorrhea, the long-term effect is stable and durable. primary dysmenorrhea of cold-damp stagnation type with acupuncture of Liji therapy has definitely long term curative effect. In addition, the treatment of patients with acupuncture of Liji therapy is more satisfactory, it is worthy of cilnlcal application.

A kinetic method for the detemination of trace rhodium based on the catalytic effect of Rh on the oxidative color reaction of diantipyry1-(p-dimethylamino)-phenylmethane (DAMAM) with potassium periodate in the H3PO4 medium and ot wateer bath was estabished. The optimum condition and kinetic parameters of this reaction were studied in detail. The linear range of the determination was 0～80μg/L, and the detection limit was 6.20×10-7g/L. The apparent activation energy og catalytic reaction was found to be 89.03kJ/mol, rate constant was 2.51×10-4/s. The system wasstable at least 6h. The method has been applid to determine trace rhodium in some catalyst samples with the relative standard deviation of 2.7%～3.2%(n=6) and recovery of 98.2%～103.2% (n=5).