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Objective To observe the effect of neuromuscular electrical stimulation on bone angiogene-sis and osteoporosis induced by denervation in sarcopenia osteoporosis rats,and explore the correlation of the miRNA expression level in circulating extracellular vesicles to bone angiogenesis and osteoporo-sis.Methods Ten-week-old male Sprague-Dawley rats were randomly divided into a sham operation control group(con,n=8),a double leg tibial nerve loss group(DD,n=8)and a double leg tibial nerve loss + right leg electrical stimulation group(D+ES,n=8).The tibial nerve of both legs was re-moved in DD and D+ES groups,and one week later the D+ES group received 8-week neuromuscular electrical stimulation.Samples were taken 24 hours after the last stimulation.Trabecular bone parame-ters were detected using micro-CT and its morphology was observed using hematoxylin and eosin(HE)staining.Immunohistochemical staining was used to identify the vascular markers platelet endothe-lial cell adhesion molecule-1(PECAM-1/CD31)and vascular endothelial growth factor A(VEGFA).Moreover,plasma exosomes were extracted and purified,and the vesicles were identified by electron microscopy,nanoparticle tracking analysis(NTA)and Western blotting.Meanwhile,the real time PCR was used to determine the expression of myogenic miR-1,miR-133a,miR-133b and miR-206 in exo-somes.Results(1)Compared with the CON group,the volume and weight of gastrocnemius and sole-us muscles in the DD group reduced significantly(P＜0.001),and the tibial trabecular bone parameters changed pathologically.After 8-week stimulation,compared with the DD group,the volume and weight of gastrocnemius and soleus muscles in D+ES group increased significantly(P＜0.05),and tibial trabecular bone parameters improved significantly.(2)The immunohistochemical staining results indicat-ed that,compared with CON group,the expression of CD31 and VEGFA in the tibia of DD group re-duced significantly,while that of the D+ES group increased significantly compared with DD group.(3)The results of NTA showed that,compared with CON group,the concentration of circulating extra-cellular vesicles in the DD group decreased significantly(P＜0.001),and the expression of miR-1,miR-133a,miR-133b and miR-206 in exosomes increased significantly(P＜0.05,P＜0.01,P＜0.05,P＜0.05 respectively).Compared with the DD group,the concentration of circulating exosomes in D+ES group increased significantly(P＜0.05),and the expression of the above four miRNAs decreased signifi-cantly(P＜0.05 for all).Conclusion Tibial nerve denervation can successfully establish the model of sar-copenia osteoporosis and neuromuscular electrical stimulation significantly relieves such denervation in-duced changes.Moreover,electrical stimulation also significantly increases the concentration of circulat-ing exosomes,and the expression of myogenic miRNAs carried by them,which suggests that the latter may be related to bone angiogenesis and the progression of osteoporosis.
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Objective To investigate the distribution of LAP+CD4+T cells in gastric carcinoma microenvironment and the correlation of LAP + CD4 + T cells with the progression of gastric tumor. Methods Forty gastric tumor patients and 20 healthy donors were enrolled in this study. The percentage of LAP+CD4+T cells in the peripheral blood and tumor tissue was detected by flow cytometry. The correlation of LAP+CD4+T cells with the progression of gastric tumor was analyzed. Results The percentage of LAP+CD4+T cells in the peripheral blood of gastric tumor patients was higher than that of health donors:(10.9±3.3)%vs. (4.3 ± 1.2)%;the percentage of LAP+CD4+T cells from tumor tissue was higher than that from non-tumor tissue:(13.5 ± 5.3)%vs. (4.7 ± 1.4)%. The percentage of LAP+CD4+T cells in peripheral blood from metastasis patients was higher than that from non-metastasis patients and health donors: (10.1 ± 6.4)% vs. (4.5 ± 1.3)% and (4.3 ± 1.2)%. Conclusions LAP+ CD4+ T cells accumulates in gastric carcinoma microenvironment and the percentage of LAP+CD4+T cells increases along with the progression of gastric tumor.
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Objective To investigate the distribution of LAP+CD4+T cells in gastric carcinoma microenvironment and the correlation of LAP + CD4 + T cells with the progression of gastric tumor. Methods Forty gastric tumor patients and 20 healthy donors were enrolled in this study. The percentage of LAP+CD4+T cells in the peripheral blood and tumor tissue was detected by flow cytometry. The correlation of LAP+CD4+T cells with the progression of gastric tumor was analyzed. Results The percentage of LAP+CD4+T cells in the peripheral blood of gastric tumor patients was higher than that of health donors:(10.9±3.3)%vs. (4.3 ± 1.2)%;the percentage of LAP+CD4+T cells from tumor tissue was higher than that from non-tumor tissue:(13.5 ± 5.3)%vs. (4.7 ± 1.4)%. The percentage of LAP+CD4+T cells in peripheral blood from metastasis patients was higher than that from non-metastasis patients and health donors: (10.1 ± 6.4)% vs. (4.5 ± 1.3)% and (4.3 ± 1.2)%. Conclusions LAP+ CD4+ T cells accumulates in gastric carcinoma microenvironment and the percentage of LAP+CD4+T cells increases along with the progression of gastric tumor.
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Objective To investigate the effects of metformin and exaliplatin on proliferation and apoptosis of gastric cancer cell line SGC-7901. The antitumor effects of metformin combined with exaliplatin on gastric cancer cell line SGC-7901 were compared. Methods SGC-7901 cells were cultured in vitro. The proliferation inhibition rate of cancer cell after treatment with various concentrations of metformin(5.0,10.0,15.0 mmol/L)、exaliplatin(2.5,5.0,10.0 μmol/L)or 15.0 mmol/L metformin combined with 10.0 μmol/L exaliplatin was analyzed via MTT colorimetric assay at 24,48,72 h.After 24 h, the SGC-7901 cell apoptosis was detected by flow cytometry, and the relative activity of caspase-3 and bax was detected by western blot. Results The co-administration of metformin and exaliplatin or single agent treatment could increase the proliferation inhibition rates of SGC-7901 cell in a time-and dose-dependent manner, while there was a significant higher proliferation inhibition rate in co-administration versus single-agent treatment (P < 0.05). The apoptosis rate induced by 15.0 mmol/L metformin and 10.0 μmol/L exaliplatin and combined treatment was (16.0 ± 2.1)%, (19.0 ± 2.7)%, (29.0 ± 3.5)%, higher than that in blank control group (10.0 ± 1.1)%, and there was significant difference (P <0.05). Both of metformin and exaliplatin could increase caspase-3 and Bax activity.Caspase-3 and Bax activity in the combination of metformin and exaliplatin was significantly higher than that of metformin group and exaliplatin group (P < 0.05). Conclusions Metformin combined with exaliplatin has synergistic effect in inhibiting the growth and inducing apoptosis of gastric cancer cell line SGC-7901.Enhancing caspase-3 and Bax expression may be the related mechanism
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AIM: To probe into the anti-epilepsy action of artificial Calculus Bovis,by observing its effect on the behavioral of the experimental epileptic rats,neuron loss in the hippocampus and hilus,and GAD positive cell alteration in the hippocampus.METHODS: SD rats were divided into three groups: group A(artificial Calculus Bovis treatment group);group B(acute epilepsy group) and group C(control group).A model of acute epilepsy rats was established by PTZ.The rat's behavioral alteration was observed by the Racine' scale.The neurons in the hippocampus and hilus were calculated by Nissl staining.The GAD positive cells were observed by immunohistochemical staining.RESULTS: The latency of the first seizure in group A was longer than that in group B,while the seizure times in group A was less than that in group B.Besides,in group A,both the neuron loss amount in the hippocampus and hilus and the GAD positive cell loss amount in the hippocampus were less than those in group B.CONCLUSION: The artificial Calculus Bovis prolonged the latency of the first seizure time,decreased the frequency of seizure,and prevented the neuron loss and protected the GAD positive cells.