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Objective To investigate the molecular mechanism of sulforaphane(Sul)promoting bone marrow stem cells(BMSCs)differentiating into osteoblasts.Methods BMSCs were divided into the control group(without any treatment),induction group(induction of osteogenic differentiation),and induction+Sul group(induction of osteogenic differentiation with the addition of 40 μmol/L of Sul).The adenovirus-shRNA-Mock,-shRNA-TET1,-shRNA-TET2,and-shRNA-TET3 were transfected into BMSCs as the shRNA-Mock group,shRNA-TET1 group,shRNA-TET2 group,and shRNA-TET3 group.BMSCs were cultured in cell culture medium containing osteogenic differentiation induction medium and 40 μmol/L of Sul,and then transfected with adenovirus-shRNA-TET1,-shRNA-TET2,-shRNA-TET3,and-shRNA-Mock as the induction+Sul+shRNA-TET1 group,induction+Sul+shRNA-TET2 group,induction+Sul+shRNA-TET3 group,and induction +Sul+shRNA-Mock group.The mRNA and protein expression levels of Runx2 after BMSCs differentiated into osteoblasts were determined by qPCR and Western blot.The DNA content of Runx2 promoter region bound to Histone H3 after BMSCs differentiated into osteoblasts was determined by chromatin immunocoprecipitation(ChIP).The methylation level of Runx2 promoter region of BMSCs differentiated into osteoblasts was determined by HpaⅡenzyme and MspⅠenzyme digestion combined with qPCR.The degree of BMSCs differentiated into osteoblasts was determined by alizarin red staining.Results Compared with the induction group,the mRNA and protein expression levels of Runx2 in the induction+Sul group were significantly increased(P<0.05);the content of DNA in the Runx2 promoter region bound to Histone H3 was increased(P<0.05),the methylation level of Runx2 promoter region was reduced(P<0.05),and the alizarin red staining score was elevated(P<0.05).Compared with the induction+Sul group,the content of DNA in the Runx2 promoter region bound to Histone H3 in the induction+Sul+shRNA-TET1 group was decreased(P<0.05),the methylation level of Runx2 promoter region was increased(P<0.05),and the alizarin red staining score was decreased(P<0.05).While there was no significant change among the induction+Sul+shRNA-TET2 group,induction+Sul+shRNA-TET3 group,induction+Sul+shRNA-Mock group(P>0.05).Conclusion Sul can promote the differentiation of BMSCs into osteoblasts through promoting DNA demethylation of Runx2 promoter region by TET1.
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Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.
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Objective: To analyze the characteristics of heart rate variability (HRV) in patients with vestibular migraine (VM) and to explore its possible mechanism. Methods: Forty-eight patients with VM [17 males and 31 females, age (36.2±9.2) years], 44 patients with migraine [15 males and 29 females, age (34.4±9.0) years], and 30 patients with health check-ups during the same period [12 males and 18 females, age (34.6±6.5) years old] were selected as study subjects. Ambulatory ECG monitoring was performed in all subjects, and the HRV characteristics of each group were analyzed from both daytime and nighttime time phases. Time domain parameters were analyzed: standard deviation of normal to normal (SDNN), root mean square of successive differences (RMSSD), and percentage of normal to normal intervals differing by more than 50 ms (pNN50). The parameters in the frequency domain were analyzed: high frequency power (HF), low frequency power (LF), and the ratio of low frequency to high frequency power (LF/HF). Statistical analysis of the data was performed using SPSS 26.0 software. Results: At night, RMSSD (F=6.694) and HF (F=9.434) were lower in the VM and migraine groups compared to the control group, while LF/HF (F=16.049) and LF (F=9.434) were elevated compared to the control group, with statistically significant differences (P<0.05 or P<0.01), while LF was significantly elevated in the VM group compared to the migraine group, with a statistically significant (P<0.05). On the daytime measurements, mainly LF was elevated in the vestibular migraine group compared with the control group, while RMSSD was decreased compared with the control group, with statistically significant differences (P<0.05). Conclusion: Autonomic dysfunction characterized by sympathetic hyperfunction and vagal hypofunction is present in VM patients and is more pronounced at night. In addition, the degree of autonomic dysfunction may be more pronounced in VM patients than in migraine patients.
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Adult , Female , Humans , Male , Middle Aged , Heart Rate/physiology , Migraine Disorders , VertigoABSTRACT
The long-term and extensive use of chemical fertilizers and pesticides in the cultivation of Chinese materia medica has resulted in serious soil ecological and environmental problems such as secondary salinization, soil consolidation, soil acidification, continuous cropping obstacles, micro-ecological imbalance, and serious soil pests and diseases in the production areas of Chinese materia medica. Therefore, promoting the ecological planting of Chinese materia medica is the only way for the production of Chinese materia medica. Attapulgite(ATP) is a kind of water-rich magnesium-rich aluminosilicate clay mineral with layered and chain structure. It has abundant reserves in China, possesses nano-material properties, strong adsorption and ion exchange properties, and has huge high value utilization space. ATP and its functional products have the potential of water and fertilizer conservation, regulating soil structure and micro-ecology, and are widely used in ecological planting of Chinese materia medica. This paper reviews the resource distribution, structural characteristics, the research and application progress in soil ecological effects of ATP, and prospects the application prospects of it in the ecological planting of Chinese materia medica.
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China , Drugs, Chinese Herbal , Magnesium Compounds , Materia Medica , Medicine, Chinese Traditional , Silicon Compounds , SoilABSTRACT
Custom ocular prosthesis as a kind of product to make up the eye defects. It can not only improve patient's comfort adaptation and restore the normal structures but also let the patient regain confidence. However, it is in infant stage in our country for custom ocular prosthesis that the professional engineer is not enough, standards for the products are shortage. These factors together result in poor understanding of product management control and impede the development of ocular prostheses. This article gives the suggestions for quality management mainly focus on human resources, raw material, production and so on.
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Objective To study the local irritation of repeated intrathecal injection of Ziconotide Acetate,and to provide reference for irritancy evaluation ofintrathecal injection.Methods Sixteen New Zealand rabbits were assigned into two groupsat random:Control group and Ziconotide Acetate group,eight animals each group.Totally 50 μL saline or Ziconotide Acetate (100 μg/mL) were administrated by repeated lumbar intrathecal injection once daily for 7 d.Animal behavior was observed every day,and four animals in each group were sacrificed 2 d later after the last injection,the lumbar spinal cord was removed for histopathological examination and irritancy evaluation.The remaining animals were sacrificed for initancy evaluation 14 d later after the last injection.Results Only one animal died after anesthesia on day three in saline group,while no obvious adverse reactions were observed in other rabbits during the entire study,and no intrathecal irritant reactions of histopathological examination were found in both groups.The reversible minor mechanical damage was observed at the injection point,2 d after the last administration.Conclusion For 7 d repeated lumbar intrathecal injection in rabbits,no intrathecal irritant reactions observed in Ziconotide group,and the New Zealand rabbit could be used as a local irritation evaluation model.
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Aim To establish a cell model targeting on GLP-1 R,and evaluate its function by the cAMP assay,for screening the new class of GLP-1 analogues as anti-diabetic candidates. Methods An eukaryotic expression vector pEGFP-GLP-1 R was constructed and transfected into HEK293A cells.After selecting with G41 8,a cell line stably expressing GLP-1 R-GFP was estab-lished.The expression and the cellular distribution of GLP-1 R-GFP fusion protein were investigated by Western blot and fluo-rescence microscopy.Then,the activity of GLP-1 analogue Lira-glutide was evaluated by monitoring the content of cAMP via HTRF using this cell model.Results GLP-1 R-GFP-293A cell line was successfully established.GLP-1 R-GFP fusion proteins were mainly distributed in the cell membrane.The dose-respon-sive relationship experiments revealed that cAMP could be effec-tively stimulated by Liraglutide using this cell model.Conclu-sion This cell model could be used to detect the bioactivity of GLP-1 analogues in vitro,which lays a foundation for the screen-ing of GLP-1 analogues and small GLP-1 R agonists.
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Codon bias is an important factor which influences heterologous gene expression. Optimizing codon sequence could improve expression level of heterologous gene. In order to improve the expression level of BmK AngM1 gene encoding the analgesic peptide from Buthus martensii Karsch in Pichia pastoris, the codon-optimized BmK AngM1 gene according to its cDNA sequence and the preference codon usage of P. pastoris were cloned into expression vector pPIC9K and then transformed into P. pastoris. The expersion of recombinant BmK AngM1 (rBmK AngM1) was inducced by methanol in the medium, and the expression level of the optimized BmK AngM1 gene was 3.7 times of the native one. These results suggested that the expression of BmK AngM1 in P. pastoris could be successfully improved by codon optimization.
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Animals , Amino Acid Sequence , Codon , Genetics , Gene Expression , Pichia , Genetics , Metabolism , Plasmids , Recombinant Proteins , Genetics , Metabolism , Scorpion Venoms , Genetics , Metabolism , Scorpions , Chemistry , Transformation, GeneticABSTRACT
Codon bias is an important factor which influences heterologous gene expression. Optimizing codon sequence could improve expression level of heterologous gene. In order to improve the expression level of BmK AngM1 gene encoding the analgesic peptide from Buthus martensii Karsch in Pichia pastoris, the codon-optimized BmK AngM1 gene according to its cDNA sequence and the preference codon usage of P. pastoris were cloned into expression vector pPIC9K and then transformed into P. pastoris. The expersion of recombinant BmK AngM1 (rBmK AngM1) was inducced by methanol in the medium, and the expression level of the optimized BmK AngM1 gene was 3.7 times of the native one. These results suggested that the expression of BmK AngM1 in P. pastoris could be successfully improved by codon optimization.
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Objective: To observe the influence of electroacupuncture on depression and defects of neurological functions in the patients with post-stroke depression. Methods: 90 cases of the apoplectic patients confirmed clinically by CT or MRI were selected and divided randomly into three groups by digital table, 30 cases in each group. All three groups of the patients were treated for wind stroke, with electric stimulation added for the acupuncture group,Fluoxetine added for medication group, and placebo added for control group, continuously for 60 d. Results: Compared to the control group, the scores in Hamilton Depression Rating Scale (HAMD) and Modified Edinburgh-Scandinavia Stroke Scale (MESSS) decreased in the electroacupuncture group and medication group, and the scores in Barthel index increased (P<0.05). The general therapeutic effects were better in the electroacupuncture group and medication group than in the control group, and the general therapeutic effects were similar in the electroacupuncture group and medication group. Conclusion: Eiectroacupuncture can improve depressive behavior and promote rehabilitation of the nervous functions.
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<p><b>BACKGROUND</b>Artificial joints present certain problems such as osteal absorption and lysis induced by wear debris which leads to loosening of the prosthesis over a period of time. Here we propose a design of an artificial magnetic suspension joint that was prepared by integrating the medical theories of modern material science, magnetism, and medical physics.</p><p><b>METHODS</b>According to clinical characteristic of biological and mechanical for hip joint, we designed the appearance and dimensions of magnetic suspension joint and placed neodymium-iron-boron permanent magnets in the prosthesis. As the same time, we performed mechanical and biological experiments using artificial magnetic suspension hip joints models.</p><p><b>RESULTS</b>By simulated the human hip structure and the external load, we discovered the artificial magnetic suspension hip joints models had much lesser amount and size of wear debris than the ceramic/ceramic artificial hip joint prosthesis in friction wear tests. The force between the artificial joints with magnetic materials that we have calculated is feasible for application of artificial joint. The design of artificial magnetic suspension hip joints models was plausible technically and safe biologically.</p><p><b>CONCLUSION</b>Artificial magnetic suspension hip joints may effectively reduce the incidence of the loosening of prosthesis over a period of time.</p>
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Humans , Hip Prosthesis , Magnetics , Prosthesis Design , SuspensionsABSTRACT
[Objective]To explore the relationship between Pglycoprotein,survivin expression and chemotherapy outcome.[Method]The expressions of P-glycoprotein,and of survivin were examined by the immunohistochemistry in 38 specimens of preoperation biopsy of osteosarecomas.[Result]The response rate of the P-glycoprotein positive group was lower than that of P-glycoprotein negative group.The response rate of the survivin overexpression group was significant lower than survivin was found to be more valuable in predicting chemotherapy results.[Conclusion]The expressions of P-glycoprotein,and of survivin are clearly associated with osteosarcomas response to chemotherapy in osteosarecoma and survivin is a more valuable parameter.