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BACKGROUND@#Triple-negative breast cancer (TNBC) is a type of highly invasive breast cancer with a poor prognosis. According to new research, long noncoding RNAs (lncRNAs) play a significant role in the progression of cancer. Although the role of lncRNAs in breast cancer has been well reported, few studies have focused on TNBC. This study aimed to explore the biological function and clinical significance of forkhead box C1 promoter upstream transcript (FOXCUT) in triple-negative breast cancer.@*METHODS@#Based on a bioinformatic analysis of the cancer genome atlas (TCGA) database, we detected that the lncRNA FOXCUT was overexpressed in TNBC tissues, which was further validated in an external cohort of tissues from the General Surgery Department of the First Affiliated Hospital of Nanjing Medical University. The functions of FOXCUT in proliferation, migration, and invasion were detected in vitro or in vivo. Luciferase assays and RNA immunoprecipitation (RIP) were performed to reveal that FOXCUT acted as a competitive endogenous RNA (ceRNA) for the microRNA miR-24-3p and consequently inhibited the degradation of p38.@*RESULTS@#lncRNA FOXCUT was markedly highly expressed in breast cancer, which was associated with poor prognosis in some cases. Knockdown of FOXCUT significantly inhibited cancer growth and metastasis in vitro or in vivo. Mechanistically, FOXCUT competitively bounded to miR-24-3p to prevent the degradation of p38, which might act as an oncogene in breast cancer.@*CONCLUSION@#Collectively, this research revealed a novel FOXCUT/miR-24-3p/p38 axis that affected breast cancer progression and suggested that the lncRNA FOXCUT could be a diagnostic marker and therapeutic target for breast cancer.
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Humanos , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Regulación Neoplásica de la Expresión Génica/genética , Sistema de Señalización de MAP Quinasas , MicroARNs/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , ARN Largo no Codificante/metabolismo , Neoplasias de la Mama Triple Negativas/patologíaRESUMEN
China is one of the countries with the highest incidence of esophageal cancer, which is still increasing year by year in recent years. Surgical treatment is the first choice for early and middle esophageal cancer. Surgeons have been exploring how to remove the tumor as completely as possible and reduce the trauma as far as possible. In recent years, with the rapid development of minimally invasive surgery and endoscopic technology, minimally invasive esophagectomy(MIE)has led the trend of radical surgery for esophageal cancer. At present, the mainstream minimally invasive surgery is thoracoscopic thoracoscopic(VATS)resection of esophageal cancer, which requires thoracotomy and anesthesia, resulting in large surgical trauma and more complications of postoperative circulatory respiratory system. Mediastinose-assisted esophagectomy(MAE), which eliminates a thoracotomy, is also an important part of MIE. Overseas MAE application started early, but the domestic development is relatively slow. This article summarizes the experience of different MAE surgical methods in China, and provides the basis for its promotion in China.
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OBJECTIVE@#A rapid fixation device is developed to solve the problems of emergency fixation and transportation of patients with spinal injury.@*METHODS@#Through the analysis of the function,3D modeling design, finite element analysis and optimization design based on ANSYS Workbench, tensile strength verification experiment, we produced the prototype, and tested it, conducted a simulated rescue experiment.@*RESULTS@#The fixation device designed can meet the demand of spinal injury patients for safe rescue after accidents, and the quality of the rapid fixation device was lighten by about 30% without reducing the intensity.@*CONCLUSIONS@#The method based on optimal design can obviously improve the structure design, and has reference significance for other related rescue equipment design.
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Humanos , Diseño de Equipo , Análisis de Elementos Finitos , Resistencia a la TracciónRESUMEN
Objective:To compare the efficacy of the combination of abidol, lopinavir/ritonavir plus recombinant interferon α-2b (rIFNα-2b) and the combination of lopinavir/ritonavir plus rIFNα-2b for patients with COVID-19 in Zhejiang province.Methods:A multicenter prospective study was carried out to compare the efficacy of triple combination antiviral therapy and dual combination antiviral therapy in 15 medical institutions of Zhejiang province during January 22 to February 16, 2020. All patients were treated with rIFNα-2b (5 million U, 2 times/d) aerosol inhalation, in addition 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir/ritonavir (2 tablets, 1 time/12 h) (triple combination group) and 41 patients were treated with lopinavir/ritonavir (2 tablets, 1 time/12 h) (dual combination group). The patients who received triple combination antiviral therapy were further divided into three subgroups: <48 h, 3-5 d and >5 d according the time from the symptom onset to medication starting. The therapeutic efficacy was compared between triple combination group and dual combination group, and compared among 3 subgroups of patients receiving triple combination antiviral therapy. SPSS 17.0 software was used to analyze the data.Results:The virus nucleic acid-negative conversion time in respiratory tract specimens was (12.2±4.7) d in the triple combination group, which was shorter than that in the dual combination group [(15.0±5.0) d] ( t=6.159, P<0.01). The length of hospital stay in the triple combination group [12.0 (9.0, 17.0) d] was also shorter than that in the dual combination group [15.0 (10.0, 18.0) d] ( H=2.073, P<0.05). Compared with the antiviral treatment which was started within after the symptom onset of in the triple combination group, the time from the symptom onset to the viral negative conversion was 13.0 (10.0, 17.0), 17.0 (13.0, 22.0) and 21.0 (18.0, 24.0) d in subgroups of 48 h, 3-5 d and >5 d, respectively ( Z=32.983, P<0.01), while the time from antiviral therapy to viral negative conversion was (11.8±3.9), (13.5±5.1) and (11.2±4.3) d, respectively( Z=6.722, P<0.05). Conclusions:The triple combination antiviral therapy of abidol, lopinavir/litonavir and rIFNα-2b shows shorter viral shedding time and shorter hospitalization time, compared with the dual combination antiviral therapy; and the earlier starting triple combination antiviral therapy will result in better antiviral efficacy.
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Objective@#Comparing the benefit of Abidor, lopinavir/ritonavir and recombinant interferon α-2b triple combination antiviral therapy and lopinavir/ritonavir and interferon dual combination antiviral therapy to hospitalized novel coronavirus pneumonia 2019 in Zhejiang province.@*Methods@#A multi-center prospective study was carried out to compare the effect of triple combination antiviral therapy with dual combination antiviral therapy in 15 medical institutions of Zhejiang Province. All patients were treated with recombinant interferon α-2b (5 million U, 2 times/d) aerosol inhalation. 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir / ritonavir (2 tablets, 1 time/12 h) as the triple combination antiviral treatment group. 41 patients were treated with lopinavir / ritonavir (2 tablets, 1 time/12 h) as the dual combination antiviral treatment group. The patients who received triple combination antiviral therapy were divided into three groups: within 48 hours, 3-5 days and > 5 days after the symptom onset. To explore the therapeutic effects of triple combination antiviral drugs and dual combination antiviral drugs, as well as triple combination antiviral drugs with different antiviral initiate time. SPSS17.0 software was used to analyze the data.@*Results@#The time of virus nucleic acid turning negative was (12.2 ± 4.7) days in the triple combination antiviral drug group, which was shorter than that in the dual combination antiviral drug group [(15.0 ± 5.0) days] (t = 6.159, P < 0.01 ). The length of hospital stay [12 (9, 17) d] in the triple combination antiviral drug group was also shorter than that in the dual combination antiviral drug group [15 (10, 18) d] (H = 2.073, P < 0.05). Comparing the antiviral treatment which was started within 48 hours, 3-5 days and > 5 days after the symptom onset of triple combination antiviral drug group, the time from the symptom onset to the negative of viral shedding was 13 (10,16.8), 17 (13,22) and 21 (18-24) days respectively (Z = 32.983, P < 0.01), and the time from antiviral therapy to the negative of viral shedding was (11.8±3.9) , (13.5±5.1) and (11.2±4.3) d. The differences among the three groups were statistically significant (Z=32.983 and 6.722, P<0.01 or<0.05).@*Conclusions@#The triple combination antiviral therapy of Abidor, Lopinavir/Litonavir and recombinant interferon α-2b showed shorter viral shedding time and hospitalization time compared with the dual combination antiviral therapy. The earlier the time to initiate triple antiviral treatment, the shorter the time of virus shedding.
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OBJECTIVE:To establish the quality standard for the leaves of Nauclea officinalis. METHODS:Microscopic identi-fication and TLC methods were used for qualitative identification of N. officinalis. The moisture and ash of medicinal materials were determined. HPLC method was adopted to determine the content of strictosamide in medicinal materials. The determination was per-formed on Lichrospher C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(gradient elution)at the flow rate of 1 mL/min,injection volume was 10 μL. The detection wavelength was set at 226 nm,and the column temperature was 30 ℃. RESULTS:Microscopic identification of the leaves of N. officinalis had strong characteristics,and TLC sports were clear and well-separated without interference from negative control. The linear range of strictosamide were 0.0105-0.21 mg/mL(r=0.9995);RSDs of precision,stability and reproducibility tests were all lower than 2.0%. Average recoveries were 96.25-101.82%(RSD=1.86%,n=9). Total ash of medicinal materials was 5.27%-6.44%,acid insoluble ash was 0.23%-0.36%,water content was 9.48%-11.46%,strictosamide was 0.124%-1.003%. CONCLUSIONS:Established standard can be used for quality evaluation of N. officinalis.
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Objective To explore the medical care and evacuation equipment at sea in China.Methods The present situation of the medical care and evacuation equipment at sea in China was discussed from the aspects of equipment system,medical service support,mechanism for utilization,management and maintenance as well as informatization.The problems were analyzed in equipment system,support ability,equipment integration and update as well as equipment performances.Results Some measures were put forward from the aspects of equipment system,support ability,equipment integration and update as well as equipment performances.Conclusion The development of medical care and evacuation equipment at sea has to take considerations on medical service requirements at sea,integrated civilian and military uses,personnel,innovation and etc.
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This study was aimed to establish an HPLC method for the content determination ofepimedin A, epimedinB,epimedinC,epimedium glycoside,baohuosideI in epimedium flavonoids capsule. The elusion was performed on an Eclipse Plus C18column (250 mm× 4.6 mm, 5μm). The mobile phase was composed of acetonitrile and water with a gradient elution. The flow rate was set at 1 mL·min-1. The detection wave length was set at 270 nm. The column temperature was 25℃. The results showed that the linear ranges of epimedin A,epimedinB,epimedinC,epimedium glycoside,baohuosideI were 3.10-62.00μg·mL-1, 5.70-114.00μg·mL-1, 9.14-182.80μg·mL-1, 15.20-304.00μg·mL-1, and 1.56-31.20μg·mL-1, respectively. The correlation coefficientr was more than 0.999 3. The average recoveries were 101.06% (RSD = 1.05%,n = 6), 100.78% (RSD = 1.08%,n = 6), 99.17% (RSD = 1.14%,n = 6), 100.23% (RSD = 0.68%,n = 6), and 99.09% (RSD = 1.30%,n = 6), respectively. This experiment was precise, reproducible and stable. It was concluded that the method was simple and accurate, which provided a certain reference value for the multi-component assaying of epimedium flavonoids capsule.
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Objective To study the differential expression of apoptosis related genes in GJB2 gene conditional knockout mice (cCx26Pax2Cre ) cochlea membranous labyrinth ,and to explore the mechanism of GJB2 gene mutations causing deafness .Methods Two developmental stages of P10 and P18 were selected from the knock out mice and the wild type ones (BALB/C) .The total RNA was isolated from cochlear membranous labyrinth and PCR array was performed using mouse apoptosis PCR arrays .Results Compared with wild -type mice ,significant up or down -regulation in gene expression was detected in 16 genes in cCx26Pax2Cre ones at P10 .Of the 16 genes ,14 ones were down-regulated .Among the 14 genes ,9 ones can be classified as anti -apoptosis or pro -proliferation genes ,5 ones can be classified as pro -apoptosis or pro -inflammation genes .The other 2 genes expression was up-regula-ted ,and their main role was to promote apoptosis .Compared with time -matched controls ,significant up or down-regulation in gene expression was detected in 4 genes in cCx26Pax2Cre mice at P18 .Of the 4 genes ,3 ones expression was down-regulated ,were anti-apoptosis ones .The expression of the other one gene among the 4 ones was up-regulated ,which acted as pro -apoptosis genes .Bcl2l10 and Tnfrsf10b expression showed significant down or up -regulation at both stages . Compared with P10 , the expression of caspase - 8 was up - ragulated at P18 in cCx26Pax2Cre mice .Conclusion It is suggested that GJB2 mutation may up -regulate the expression of DR5 ,which can trigger the death receptor pathway to cause apoptosis in cCx 26Pax2Cre mice cochlea directly .At the same time , caspase-8 in death receptor pathway may activate the mitochondria pathway indirectly and amplify the apoptosis further in cCx26Pax2Cre mice cochlea .The final result of the above activated pathways is the wide range of cochlear cells apoptosis and the profound hearing loss in cCx 26Pax2Cre mice .
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Objective To observe the effect of mouse nerve growth factor (mNGF) on cognitive function and activities of daily living (ADL) in patients with delayed encephalopathy after acute carbon monoxide poisoning. Methods 35 patients with delayed encephalopathy after acute carbon monoxide poisoning were randomly divided into treatment group (n=18) and control group (n=18). Both groups accepted hyperbaric oxygen and medication, while the treatment group was injected with mNGF 20 mg a day in addition for 12 weeks. They were as-sessed with Loewenstein Occupational Therapy Cognitive Asessment (LOTCA) and modified Barthel Index (MBI) before and after treat-ment. Results The scores of LOTCA and MBI improved more in the treatment group than in the control group after treatment (P<0.05). Conclusion mNGF can obviously improve cognitive function and ADL for patients with delayed encephalopathy after acute carbon monox-ide poisoning.
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BACKGROUND:A preliminary experiment developed a double-chamber stirred bioreactor which can carry out osteogenic and cartilage induction at the same time. OBJECTIVE:To explore the effects of double-chamber stirred bioreactor on the repair of goat knee cartilage defects with tissue-engineered cartilage. METHODS:Twelve goats were selected to make bilateral femoral condyle osteochondral defects models and randomized to three groups:experimental group, implanted with the composites ofβ-tricalcium phosphate and bone marrow mesenchymal stem cells that were subjected to 2-week chondrogenic and osteogenic induction simultaneously in the double-chamber stirred bioreactor under mechanical stimulation;control group, implanted with the composites ofβ-tricalcium phosphate and bone marrow mesenchymal stem cells that were subjected to 2-week chondrogenic and osteogenic induction simultaneously in the double-chamber stirred bioreactor;blank control group, without treatment. After 12 and 24 weeks of implantation, general observation, Masson staining, II col agen immunohistochemical staining and histological scoring were performed. RESULTS AND CONCLUSION:In the experimental and control groups, new cartilage tissue and bone tissue were visible, but the experimental group showed better repair effects than the control group (P<0.05). The blank control group had no cartilage formation. These findings indicate that under the mechanical stimulation by the double-chamber stirred bioreactor in vitro, the repair effect of tissue-engineered osteochondral complex on knee joint cartilage defects can be improved.
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Objective To observe the effect of apigenin on acute lung injury (ALI) induced by lipopolysaccharide(LPS)in mice,and to discuss its possible mechanism. Methods Forty healthy male Kunming mice were randomly divided using random number table into control group,model group and low,medium,high dose groups of apigenin intervention,and each group consisted of 8 mice. The model of ALI was reproduced by intraperitoneal injection of 5 mg/kg LPS. Mice of the low,medium and high-dose intervention groups were given intraperitoneal injection of apigenin 10,25,50 mg/kg,respectively,1 hour before LPS modeling. Pathological changes in right upper lobe of lung tissue were examined after hematoxylin and eosin(HE)staining and pathology score was observed at 6 hours after modeling. Right inferior lung was weighed to measured wet/dry ratio(W/D). Intercellular adhesion molecule-1(ICAM-1)and tumor necrosis factor-α(TNF-α)in serum and bronchoalveolar lavage fluid (BALF)were determined by enzyme linked immunosorbent assay(ELISA). The mRNA expressions of p38 mitogen-activated protein kinase(p38MAPK),ICAM-1,and TNF-α were determined by reverse transcription-polymerase chain reaction(RT-PCR). Results Compared with control group,lung W/D ratio in model group was significantly increased(17.79±2.89 vs. 5.56±0.37,P<0.05),and the pathology score was significantly elevated(10.32±0.23 vs. 1.87±0.54,P<0.05),ICAM-1 and TNF-α contents,in serum and BALF were increased〔ICAM-1(ng/L) in serum:21.4±2.7 vs. 14.3±3.5,TNF-α(ng/L)in serum:254.8±10.6 vs. 142.3±13.7;ICAM-1(ng/L)in BALF:20.3±2.4 vs. 11.5±3.2,TNF-α(ng/L)in BALF:230.3±5.8 vs. 110.5±11.2,all P<0.05〕,and the mRNA expressions of p38MAPK,ICAM-1 and TNF-α were also increased significantly(the mRNA expression of p38MAPK,ICAM-1 and TNF-αwere 4.42±0.37,4.89±0.27,3.28±0.13,respectively,all P<0.05). Different doses of apigenin could obviously alleviate the damaging effect to the lung,and the most obvious effect was seen in the medium dose group,in which lung W/D ratio was 13.28±1.21,ICAM-1 in serum was(18.5±4.3)ng/L,TNF-αin serum was(169.4±20.8)ng/L,ICAM-1 in BALF was(17.8±3.5)ng/L,TNF-αin BALF was(150.4±7.1)ng/L, the mRNA expression of p38MAPK,ICAM-1 and TNF-αin lung tissue was 2.99±0.28,3.97±0.17,2.87±0.27, respectively. Statistically significant difference was found when they were compared with that of model group(P<0.05 or P<0.01). Conclusion Different doses of apigenin have some antagonistic effect against LPS in producing ALI in mice,the best improvement effect was seen in the medium dose group,and the protective effect may be related to inhibition of p38MAPK signaling pathway activity and reduction of pro-inflammatory factors such as TNF-αand ICAM-1 expression.
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@#Objective To observe the effect of mouse nerve growth factor (mNGF) on cognitive function and activities of daily living (ADL) in patients with delayed encephalopathy after acute carbon monoxide poisoning. Methods 35 patients with delayed encephalopathy after acute carbon monoxide poisoning were randomly divided into treatment group (n=18) and control group (n=18). Both groups accepted hyperbaric oxygen and medication, while the treatment group was injected with mNGF 20 mg a day in addition for 12 weeks. They were assessed with Loewenstein Occupational Therapy Cognitive Asessment (LOTCA) and modified Barthel Index (MBI) before and after treatment. Results The scores of LOTCA and MBI improved more in the treatment group than in the control group after treatment (P<0.05). Conclusion mNGF can obviously improve cognitive function and ADL for patients with delayed encephalopathy after acute carbon monoxide poisoning.
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Objective To explore the methods of breeding ,reproduction and genotype of GJB2 knock -out (cCx26KO) mice and further study the critical role of GJB2 mutation in the onset of nonsyndromic hearing loss (NSHL) .Methods Two pairs of transgenic mice (Cx26loxp/loxp and Pax2 -Cre/+ ) were inbreeded to produce Cx26loxp/-_Pax2-Cre/+ ones ,female of which were used to mate with the male Cx26loxp/loxp ones to finally get the Cx26loxp/loxp_Pax2 -Cre/+ mice(cCx26KO) .The genotype was done by PCR and Agarose gel electro-phoresis using genome DNA extracted from the mice tails .The c-ABR was used to detect the hearing ability of the cCx26KO mice .Results Both breeding and reproduction of cCx26KO mice were successful .It was fruitful to obtain four genotype mice(Cx26loxp/loxp_ Pax2-Cre / + ,Cx26loxp / -_Pax2-Cre / + ,Cx26loxp/loxp ,Cx26loxp /-) by the breeding Cx26loxp / -_Pax2Cre / + and Cx26loxp/loxp mice .The results of breeding were met with the Mendel's law .The c-ABR revealed elevated response threshold around 95 dB SPL in cCx26KO mouse compared to the wild type ones ,which further validated the accuracy of the PCR method .Conclusion The PCR method is cor-rectly identified sub pups genotype and the female Cx26loxp/-_Pax2-Cre/+ mice mating with the male Cx26loxp/loxp ones is an effective way to obtain the cCx 26KO mice .
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Objective To investigate the mechanism of PBEF in pulmonary vascular endothelial permeability increasing after car-diopulmonory bypass ,in order to provide the basis for a better lung protective measures during cardiopulmonary bypass .Methods Animal models were established ,group A :the rats were transfected with the lentiviral AD-PBEFshRNA ;group B :the rats were took 30 min deep hypothermic circulatory arrest ;group C :the rats were took 30 min deep hypothermic circulatory arrest ,then trans-fected with the lentiviral AD-PBEFshRNA ;control group :the rats were anesthetized and established CPB tube ,without CPB by-pass .Lung tissue was detected with Western blotting and ELISA .Results PBEF ,phosphorylation of P38MAPK ,ERK ,MLC ,VE-cadherin ,FAK in group C had significant difference with group A ,B and the control group(P<0 .05) .VEGF ,MMP2 ,MMP9 ,W/D in group C had significant difference with group A ,B and the control group(P<0 .05) .The pathological results showed that rat lung tissue of the control group was normal ,while in group C ,it had severe pathological damage ,the pathological damage degree in group A ,B reduced compared to group C .Conclusion PBEF can through MAPK/PI3K-Akt/VEGF signaling pathway ,increase pulmonary vascular endothelial permeability .
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OBJECTIVES: Apoptosis may play an important role in the mechanism underlying the GJB2 gene conditional knockout (cCx26) mice cochlear cell death. The objective of this study was to explore the the damage mode of the outer hair cells (OHCs) and its real time point of apoptosis and provide information to further explore the role of apoptosis in the happening of hearing loss in cCx26 mice. METHODS: Cochleae from mice at various developmental stages (P8, P12, and P21) were dissected out and first used to be observed under the scanning electron microscope (SEM). Basilar membranes from mice at P8, P14, P18, and P21 were stained by fluorescein isothiocyanate-conjugated phalloidin and propidium iodide (PI) and examined under confocal microscope. RESULTS: The loss of OHCs of cCx26 knockout mice was first set between P12 and P21 under SEM. Whole mount phalloidin and PI staining revealed that obvious apoptotic appearance of the OHCs surface morphology was observed at P18. CONCLUSION: Typical apoptotic morphology was found in the OHCs in the organ of Corti of the cCx26 mice at P18. This may provide information to further study the role of apoptosis in the occurrence of hearing loss of cCx26 mice.
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Animales , Ratones , Apoptosis , Membrana Basilar , Muerte Celular , Cóclea , Conexinas , Electrones , Fluoresceína , Cabello , Células Ciliadas Auditivas Externas , Pérdida Auditiva , Pérdida Auditiva Sensorineural , Ratones Noqueados , Órgano Espiral , Faloidina , PropidioRESUMEN
Objective To establish three-dimensional (3D) simulation model of acetabular fracture based on Letournel-Judet classification and analyze its significance. Methods Pelvis of one healthy female volunteer was scanned with 16 slice spiral CT. Dicom format data were processed by software Mimics 10.01 to reconstruct 3D model of hip bone. Different fracture types were simulated based on Letournel-Judet classification of acetabular fracture. The fracture fragments were displayed with different color. Screen-captures of fracture model from different directions were saved and video mode of fracture model was exported. Five orthopedic doctors and 10 medical students were employed to compare 3D fracture model with two-dimensional planar schematic diagram and made preliminary evaluation. Results The 3D simulation model of acetabular fracture had realistic form and stereoscopic sense, with satisfactory visual effect. The fracture could be observed and charted from optional direction and angle. Fracture model could rotate for 360° in the corresponding video mode. Four orthopedic doctors and nine medical students considered that 3D simulation model of acetabular fracture was helpful for understanding fracture classification. Conclusion The 3D simulation model of acetabular fracture is intuitive, realistic and dynamic and is helpful for clinical work and medical teaching.
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Objective To explore the feasibility of labeling in vitro rabbit osteoblasts with Qtracker and the features of Qtracker-labeled rabbit osteoblasts. Methods A healthy male rabbit, 3 months old, weighing 2 kg, was used in this study. After bone marrow was aspirated, bone marrow stromal cells (BMSCs) were isolated and cultured using the adherence method in vitro. The third passage of BMSCs was induced into osteablasts before incubation with Qtracker at concentrations of 1, 2, 4, 8, 16, 32 nmol/10~6 cells (Groups A, B, C, D, E, F respectively). Cells not labeled by Qtracker served as negative control (Group G). The following parameters were measured: induction, differentiation and determination of rabbit osteoblasts; the optimal mass concentration of Qtracker labeling by fluorescence microscopy and flow cytometry; the cell sur-vival rates at various concentrations of Qtraeker labeling by trypan-blue exclusion; Qtracker-labeled cell pro-liferation by MTr. Results The primary and the passage rabbit BMSCs were chiefly of fusiform shape. Rabbit BMSCs differentiated into osteoblasts following induction. The osteoblasts cytoplasm showed green fluorescence under fluorescence microscopy after being labeled by Qtracker. The mean labeling rate increased with the increased concentration of Qtracker, reaching up to (93.58±2.08) % after incubation at 8 nmol/ 10~6 cells by fluorescence microscopy, and (95.24±1.31) % by flow cytometry. There were no significant differences between Groups D, E, F(P>0.05), but significant differences were found between Groups A, B, C and Groups D, E, F (P<0.05). The labeling rate for Group G was 0. The cell survival rates were all above 96% (P>0.05) . No significant differences were found in the cell proliferation among various con-centrations (P>0.05). Conclusions Qtraeker can be used as a labeling marker for rabbit osteoblasts. When the concentration is at 8 nmol/10~6 cells, optimal labeling effect can be achieved. Rabbit osteoblasts labeled with Qtracker are of high efficiency and safety.
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Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.
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OBJECTIVE: To investigate the effects of Qufeng Tongluo Recipe (QFTLR), a compound of traditional Chinese herbal medicine for dispelling wind and removing obstruction in the meridians, on cell proliferation and expressions of transforming growth factor-beta1 (TGF-beta1) and interleukin-6 (IL-6) mRNAs induced by lippolysaccharide in glomerular mesangial cells from rats. METHODS: The method of serum pharmacology was used. A total of 32 rats were divided into normal control group, untreated group, QFTLR group and positive control group which also was named Monopril (fosinopril sodium) group. Mesangial proliferative glomerulonephritis was induced by injection of antithymocyte serum in the rats except for the normal control group. Sera of the rats were obtained after corresponding interventions. Lipopolysaccharide-induced proliferation of rat mesangial cells (MCs) cultured in the respective serum-containing media was detected by the method of methyl thiazolyl tetrazolium (MTT) assay, and the expressions of TGF-beta1 and IL-6 mRNAs were analyzed by the method of reverse transcription polymerase chain reaction. RESULTS: Compared with the untreated group, QFTLR showed remarkable inhibitory function on the proliferation of the mesangial cells (P<0.05). The expressions of TGF-beta1 mRNA in mesangial cells were increased in the untreated group, QFTLR group and Monopril group when compared with the normal control group (P<0.01), but the TGF-beta1 mRNA expressions in QFTLR group and in Monopril group were lower than that in the untreated group. The IL-6 mRNA expression could be increased by the LPS stimulation, and it was significantly higher in the other three groups than that in the normal control group, including the untreated group, the Monopril group and the QFTLR group (P<0.01). Compared with the untreated group, the expression of IL-6 mRNA was decreased by QFTLR and Monopril (P<0.01). QFTLR was better than Monopril in inhibiting the proliferation of the mesangial cells and decreasing the expressions of TGF-beta1 and IL-6 mRNAs (P<0.05). CONCLUSION: QFTLR has great inhibitory effect on mesangial cell proliferation and expressions of TGF-beta1 and IL-6 mRNAs, which may be one of its mechanisms in postponing glomerular sclerosis.