Objective:To observe the effect of low-intensity pulsed ultrasound (LIPUS) at different intensities on the expression of adiponectin and its receptors in C2C12 myoblasts, and to explore the potential mechanism by which LIPUS promotes the differentiation of C2C12 myoblasts.Methods:C2C12 myoblasts cultured in vitro were randomly divided into a control group and U 0.1, U 0.3 and U 0.5 groups. The control group received sham LIPUS exposure, while the U 0.1, U 0.3 and U 0.5 groups were exposed to LIPUS at intensities of 0.1W/cm 2, 0.3W/cm 2 or 0.5W/cm 2 respectively, and 1MHz for 5 min daily for 5 days. Cell viability was measured using CCK-8 assays. Fluorescence quantitative reverse transcription-polymerase chain reactions were used to detect the mRNA expression of adiponectin, adiponectin receptor 1 (adipoR1) and T-cadherin in the cells. Western blotting was employed to assess the protein expression of adiponectin, adipoR1, T-cadherin, adenosine monophosphate activated protein kinase (AMPK), activated phosphorylated adenylate-activated protein kinase (P-AMPK), embryonic myosin heavy chain (eMHC) and myogenin (MYOG). The differentiation ability of the 4 groups was measured using cell immunofluorescence chemistry. Results:After the intervention the cell viability in the U 0.1, U 0.3 and U 0.5 groups was significantly higher than in the control group. Compared with the control group, the average mRNA expression of adiponectin and the receptors of adipoR1 and T-cadherin were up-regulated significantly in the U 0.3 and U 0.5 groups. The average adiponectin, adipoR1 and T-cadherin protein expressions, and the AMPK phosphorylation level in the U 0.3 and U 0.5 groups had increased significantly compared with the control group, but all were significantly lower than in the U 0.3 group. The average protein expression of eMHC and MYOG, and the C2C12 myoblast fusion indices of the U 0.3 and U 0.5 groups were significantly higher the control group′s averages. Conclusions:LIPUS can promote the differentiation of C2C12 myoblasts. It is most effective at 0.3W/cm 2, administered for 5min/d at 1MHz with a 20% duty cycle. Its regulatory mechanism may be related to up-regulation of the expression of adiponectin, the adipoR1 and T-cadherin receptors, and the activation of AMPK phosphorylation in C2C12 myoblasts.

Objective To investigate the inhibitory effect of rhIFN-γ on human chronic myeloid leukemia cell line K562,and the impaction on the expression of CD123.Methods MTT method was used to test cell relative viability with rhIFN-γ at different concentrations.The expression of CD123 on K562 ceils was detected by flow cytometry.Results The relative inhibition of K562 cells proliferation was hampered when the cells were treated with rhIFN-γ for 72 h at the concentration of 5 x 10P ～ 108 U/L,respectively.However,rhIFN-γ at 2 x 105 U/L was benefit to K562 cells proliferation.After treatment with rhIFN-γ at 0,2 x 105 U/L and 107 U/L,the percentages of CD123 expression on K562 cells were (4.10 ± 1.46) ％,(7.2 ± 2.50) ％ as well as (21.6 ± 1.17) ％,respectively.Compared with the control group,107 U/L rhIFN-γ significantly increased the expression of CD123 on K562 cells (P＜0.05).Conclusion The effect of rhIFN-γ on the growth of K562 cells has two aspects (inhibition or proliferation),and it can increase the expression of CD 123.

Objective To investigate the changes of IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γexpressions in bone marrow of chro‐nic myeloid leukemia(CML)patients .Methods The IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γexpression levels were detected by u‐sing flow cytometry in 30 cases of CML chronic phase(CML‐CP) ,21 cases of CML accelerated phase(CML‐AP) ,15 cases of CML blastic phase(CML‐BP) ,42 cases of CML remission after treatment and 7 cases of non‐remission .Then the detection results were compared with those in the control group .Results The expression levels of INF‐γ and IL‐2 in each CML groups were lower than those in the control group(P<0 .05) ,while the expression levels of IL‐1β,IL‐4 ,IL‐6 and IL‐10 were higher than those in the con‐trol group(P<0 .05) .With the disease condition progression ,the INF‐γand IL‐2 levels were gradually decreased ,i .e .,CML‐BPCML‐AP> CML‐CP ,the difference among groups had statistical significance (P<0 .05) .After complete remission by treatment ,the INF‐γand IL‐2 levels were risen to some extent ,but which were still lower than those in the control group(P<0 .01) ,the expression levels of IL‐1β,IL‐4 ,IL‐6 and IL‐10 were decreased to some extent ,but likewise were higher than those in CML‐CP(P< 0 .05);the levels of IL‐1β,IL‐4 ,IL‐6 ,IL‐10 ,IL‐2 and INF‐γ had no statistical difference between the non‐remission group and CML‐BP group(P>0 .05) .Conclusion The changes of serum cytokines in bone marrow microenvironment of CMLpatients have certain significance to the occurrence ,development and prognosis of CML ;the de‐tection of IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γlevels in bone marrow is hopeful to provide new ideas and theoretical basis for im‐mune therapy and prognosis judgment of CML patients .

Objective:To explore the role of microRNA-133a (miR-133a) and silent mating information regulation 2 homolog 1 (SIRT1) in the effects of electroacupuncture on persons with disuse muscular atrophy.Methods:Thirty C57BL/6 mice were randomly divided into a control group, an experimental control group and an experimental group, each of 10. Disuse muscular atrophy was induced in the mice of the experimental and experimental control groups using tail suspension. The mice in the electroacupuncture group were given 15 minutes of electroacupuncture over the Yanglingquan and Zusanli points every day for 14 days. Wet weight ratio and the cross-sectional area of the gastrocnemius and soleus were tracked, and the structure of the mitochondria in the skeletal muscles was observed using a transmission electron microscope. The protein expressions of SIRT1, peroxisome proliferator-activated receptor γ coactivator-1a (PGC-1a), nicotinamide phosphoribosyl transferase (NAMPT), adenosine 5-monophosphate-activated protein kinase-a (AMPK-a) and phospho-AMPK-a (P-AMPK-a) were detected using western blotting. The expressions of the muscle atrophy F-box (Atrogin-1), muscle ring finger1 (MuRF1), miR-133a, SIRT1, paired box gene 7 (Pax7), myogenic determination (MyoD) and myogenin (MyoG) genes were detected through polymerase chain reactions. The concentration of Niacylamide adenine dinucleotide (NAD+ ) and the ratio of NAD+ to reduced nicotinamide adenine dinucleotide were also measured.Results:Compared with the experimental control group, the average wet weight increased by 21% and the cross-sectional area of the soleus increased by 30% in the experimental group. The average wet weight of the gastrocnemius increased by 5% and the area by 17%. The average expressions of Atrogin-1, MuRF1, SIRT1, PGC-1a and NAMPT, the concentration of NAD+ , as well as the average value of P-AMPK-a/AMPK-a and NAD+ /NADH were significantly lower in the experimental group than in the experimental control group, while the average expression of miR-133a in the experimental group was significantly (163%) higher. The average expressions of Pax7 and MyoD were significantly up-regulated in the experimental control group compared with the other two groups, while MyoG was highly expressed in the experimental group compared with the other 2 groups.Conclusions:The SIRT1 pathway is one of the reflexive protective mechanisms that mediate in the natural recovery of skeletal muscles. Electroacupuncture enhances myoblast differentiation, improves energy metabolism in the mitochondria, and thus promotes recovery from disuse muscular atrophy.

Objective:To investigate the expression of macrophage migration inhibitory factor (MIF) in bone marrow fluid and peripheral blood of patients with acute myeloid leukemia (AML) and its effect on the expression of interleukin-8 (IL-8) in bone marrow mesenchymal stem cells (BM-MSC).Methods:Fifty bone marrow fluid samples and 50 peripheral blood samples were collected from 50 patients with AML diagnosed in the First People's Hospital of Yunnan Province from October 2017 to January 2019, of which 17 patients were newly diagnosed, 26 patients were complete remission (CR), and 7 patients were partial remission (PR) or non-remission (NR). Fifty plasma samples from 50 healthy subjects and 50 bone marrow fluid samples from 50 patients with iron deficiency anemia were used as the controls. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of MIF protein in the samples, and the relationship between MIF expression level and clinicopathological characteristics of AML patients was analyzed. BM-MSC was successfully isolated and cultured from 42 bone marrow fluid samples of AML patients, the suitable samples for experiment were chosen and divided into BM-MSC control group (untreated BM-MSC), recombinant human macrophage migration inhibitory factor (rhMIF) group and rhMIF+ISO-1 group. ELISA and real-time fluorescence quantitative polymerase chain reaction were used to detect the expression level of IL-8 protein and mRNA in each BM-MSC group.Results:The expression levels of MIF protein in bone marrow fluid and plasma in AML group were (24.9±7.7) ng/ml and (60.5±12.1) ng/ml, the difference was statistically significant ( P < 0.01), and those in control group were (5.3±2.6) ng/ml and (2.0±1.1) ng/ml, respectively, and there were statistical differences between the two groups (t values were 136.71, 33.97 and 17.58, all P < 0.01). MIF protein expression levels in bone marrow fluid and plasma of AML patients in newly diagnosed group and PR+NR group were higher than those in CR group, and the differences were statistically significant (all P < 0.01). MIF protein expression levels were higher in bone marrow fluid and plasma of patients with ≥60 years of age, peripheral blood white blood cell count ≥30×10 9/L and bone marrow myeloblast ratio > 0.50 (all P < 0.05), but the differences were not statistically significant between patients with different gender (both P > 0.05). The detection results of each BM-MSC group showed that rhMIF promoted the IL-8 expression in BM-MSC at the gene and protein levels, which could be inhibited by the MIF inhibitor ISO-1 (all P < 0.01). Conclusion:The increased expression levels of MIF in bone marrow fluid and plasma of patients with AML are associated with the disease progression, and rhMIF can promote the expression of IL-8 in BM-MSC.