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1.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 46(3): 307-315, 2024 Jun.
Article in Zh | MEDLINE | ID: mdl-38548389

ABSTRACT

Objective To investigate the effects of platelet-rich plasma-derived exosomes (PRP-Exos) on the proliferation and migration of tendon stem/progenitor cell (TSPC).Methods PRP-Exos were extracted through the combination of polymer-based precipitation and ultracentrifugation.The morphology,concentration,and particle size of PRP-Exos were identified by transmission electron microscopy and nanoparticle tracking analysis.The expression levels of surface marker proteins on PRP-Exos and platelet membrane glycoproteins were determined by Western blot analysis.Rat TSPC was extracted and cultured,and the expression of surface marker molecules on TSPC was detected using flow cytometry and immunofluorescence staining.The proliferation of TSPC influenced by PRP-Exos was evaluated using CCK-8 assay and EdU assay.The effect of PRP-Exos on the migration of TSPC was evaluated by cell scratch assay and Transwell assay.Results The extracted PRP-Exos exhibit typical saucer-like structures,with a concentration of 4.9×1011 particles/mL,an average particle size of (132.2±56.8) nm,and surface expression of CD9,CD63 and CD41.The extracted TSPC expressed the CD44 protein.PRP-Exos can be taken up by TSPC,and after co-cultured for 48 h,concentrations of 50 and 100 µg/mL of PRP-Exos significantly promoted the proliferation of TSPC (both P<0.001),with no statistical difference between the two concentrations (P=0.283).Additionally,after co-cultured for 24 h,50 µg/mL of PRP-Exos significantly promoted the migration of TSPC (P<0.001).Conclusion Under in vitro culture conditions,PRP-Exos significantly promote the proliferation and migration of rat TSPC.


Subject(s)
Cell Movement , Cell Proliferation , Exosomes , Platelet-Rich Plasma , Stem Cells , Tendons , Exosomes/metabolism , Platelet-Rich Plasma/metabolism , Rats , Stem Cells/cytology , Stem Cells/metabolism , Animals , Tendons/cytology , Tendons/metabolism , Cells, Cultured , Rats, Sprague-Dawley , Male
2.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 45(3): 374-381, 2023 Jun.
Article in Zh | MEDLINE | ID: mdl-37106519

ABSTRACT

Objective To investigate the effect of human platelet-rich plasma-derived exosomes(PRP-exos)on the proliferation of Schwann cell(SC)cultured in vitro. Methods PRP-exos were extracted by polymerization-precipitation combined with ultracentrifugation.The morphology of PRP-exos was observed by transmission electron microscopy,and the concentration and particle size distribution of PRP-exos were determined by nanoparticle tracking analysis.Western blotting was employed to determine the expression of the marker proteins CD63,CD81,and CD9 on exosome surface and the platelet membrane glycoprotein CD41.The SCs of rats were isolated and cultured,and the expression of the SC marker S100ß was detected by immunofluorescence staining.The fluorescently labeled PRP-exos were co-cultured with SCs in vitro for observation of their interaction.EdU assay was employed to detect the effect of PRP-exos on SC proliferation,and CCK-8 assay to detect the effects of PRP-exos at different concentrations(0,10,20,40,80,and 160 µg/ml)on SC proliferation. Results The extracted PRP-exos appeared as uniform saucer-shaped vesicles with the average particle size of(122.8±38.7)nm and the concentration of 3.5×1012 particles/ml.CD63,CD81,CD9,and CD41 were highly expressed on PRP-exos surface(P<0.001,P=0.025,P=0.004,and P=0.032).The isolated SCs expressed S100ß,and PRP-exos could be taken up by SCs.PRP-exos of 40,80,and 160 µg/ml promoted the proliferation of SCs,and that of 40 µg/ml showed the best performance(all P<0.01). Conclusions High concentrations of PRP-exos can be extracted from PRP.PRP-exos can be taken up by SCs and promote the proliferation of SCs cultured in vitro.


Subject(s)
Exosomes , Platelet-Rich Plasma , Humans , Rats , Animals , Exosomes/metabolism , Schwann Cells , Coculture Techniques , Cell Proliferation , Cells, Cultured
3.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 42(2): 190-196, 2020 Apr 28.
Article in Zh | MEDLINE | ID: mdl-32385024

ABSTRACT

Objective To explore the value of conventional ultrasound combined with shear-wave elastography in the quantitative evaluation of sciatic nerve crush injury in rabbit models. Methods Forty healthy male New Zealand white rabbits were randomly divided into four groups (n=10 in each group):three crush injury (CI) groups (2,4,and 8 weeks after crush) and control group (without injury). The thickness and stiffness of the crushed sciatic nerves and denervated triceps surae muscles were measured at different time points and compared with histopathologic parameters. Inter-reader variability was assessed with intraclass correlation coefficients. Results Compared with the control group,the inner diameters of the sciatic nerves significantly increased in the 2-week CI group [(1.65±0.34) mm vs. (0.97±0.15) mm,P=0.00] but recovered to the nearly normal level in the 8-week CI group [(1.12±0.18) mm vs. (0.97±0.15) mm,P=0.06];however,compared with control group [(8.75±1.02)kPa],the elastic modulus of the nerves increased significantly in all the CI groups [2-week:(14.77±2.53) kPa;4-week:(19.12±3.46) kPa;and 8-week:(28.39±5.26) kPa;all P=0.00];pathologically,massive hyperplasia of collagen fibers were found in the nerve tissues. The thickness of denervated triceps surae muscle decreased gradually,and the elastic modulus decreased 2 weeks after injury but increased gradually in the following 6 weeks;pathologically,massive hyperplasia of collagen fibers and adipocytes infiltration were visible,along with decreased muscle wet-weight ratio and muscle fiber cross-sectional area. The inter-reader agreements were good. Conclusion Conventional ultrasound combined with shear-wave elastography is feasible for the quantitative evaluation of the morphological and mechanical properties of crushed nerves and denervated muscles.


Subject(s)
Crush Injuries/diagnostic imaging , Elasticity Imaging Techniques , Sciatic Nerve/injuries , Ultrasonography , Animals , Elastic Modulus , Male , Muscle, Skeletal/innervation , Muscle, Skeletal/pathology , Rabbits , Random Allocation
4.
J Ultrasound Med ; 38(5): 1191-1200, 2019 May.
Article in English | MEDLINE | ID: mdl-30208234

ABSTRACT

OBJECTIVES: The changes in the viscoelasticity of the Achilles tendon are related to tendinopathy. Therefore, constructing a data model in the healthy population is essential to understanding the key factors affecting the viscoelasticity of the Achilles tendon. The purpose of our research was to obtain large sample data, construct a data model, and determine parameters that affect the elastic modulus of the Achilles tendon in healthy Chinese adults. METHODS: We designed a prospective multicenter clinical trial to evaluate the viscoelasticity of the Achilles tendon by using shear wave elastography. A total of 1165 healthy adult participants from 17 Chinese hospitals were recruited for the assessment. The necessary parameters (age, height, weight, and body mass index) were recorded. The elastic modulus (Young modulus) was obtained from the middle of the Achilles tendon and calculated with feet in naturally relaxed, dorsal, and plantar positions. The thickness and perimeter of the Achilles tendon were measured via cross section on the same site. A multiple linear regression was performed to find the key factors affecting the Young modulus of the Achilles tendon. RESULTS: The Young modulus of the left Achilles tendon in the natural relaxed position followed a normal distribution (P > .05) with a mean ± SD of 374.24 ± 106.12 kPa. The regression equations showed a positive correlation between the Young modulus and weight and a negative correlation between the Young modulus and the circumference or thickness of the left Achilles tendon (P < .05). CONCLUSIONS: The Young modulus of the Achilles tendon as measured by shear wave elastography is related to body weight as well as the perimeter or thickness of the tendon.


Subject(s)
Achilles Tendon/physiology , Elastic Modulus/physiology , Elasticity Imaging Techniques/methods , Achilles Tendon/diagnostic imaging , Adult , China , Female , Humans , Male , Prospective Studies , Reference Values
5.
Med Sci Monit ; 22: 1186-91, 2016 Apr 09.
Article in English | MEDLINE | ID: mdl-27072885

ABSTRACT

BACKGROUND There has been no published report assessing the mechanical properties of a repaired Achilles tendon after surgery using shear wave elastography (SWE). The aim of this study was to investigate the changes in mechanical properties of the healing Achilles tendon after surgical repair of a tendon rupture using ultrasound SWE and how these changes correlate with tendon function. MATERIAL AND METHODS Twenty-six patients who underwent surgical repair for Achilles tendon rupture were examined with ultrasound SWE coupled with a linear array transducer (4-15 MHz). The elasticity values of the repaired Achilles tendon in a longitudinal view were measured at 12, 24, and 48 weeks postoperatively. Functional outcomes were assessed with the American Orthopedic Foot and Ankle Society (AOFAS) rating system at 12, 24, and 48 weeks postoperatively. General linear regression analysis and correlation coefficients were used to analyze the relationship between elasticity and the AOFAS score. RESULTS There were significant differences with respect to the mean elasticity values and functional scores of the repaired Achilles tendon at 12, 24, and 48 weeks postoperatively (all P<0.05). Tendon function was positively correlated with the elasticity of the repaired Achilles tendon (P=0.0003). CONCLUSIONS Our findings suggest that SWE can provide biomechanical information for evaluating the mechanical properties of healing Achilles tendon and predict Achilles tendon function.


Subject(s)
Achilles Tendon/injuries , Achilles Tendon/physiopathology , Tendon Injuries/physiopathology , Wound Healing/physiology , Achilles Tendon/diagnostic imaging , Achilles Tendon/surgery , Adult , Aged , Elastic Modulus , Elasticity Imaging Techniques/methods , Female , Humans , Male , Middle Aged , Orthopedic Procedures/methods , Plastic Surgery Procedures/methods , Rupture , Tendon Injuries/diagnostic imaging , Tendon Injuries/surgery , Treatment Outcome , Ultrasonics , Ultrasonography/methods
6.
Pak J Pharm Sci ; 27(4 Suppl): 983-9, 2014 Jul.
Article in English | MEDLINE | ID: mdl-25016256

ABSTRACT

Lectins are the tools for the determination of sugar chain structure. Recently, lectin arrays have become a popular new technology; therefore, lectins with specific sugar-binding properties are required. The objective of the study was to isolate a novel lectin from Pleurotus ferulae mushrooms and characterize its various biological activities. A novel lectin was extracted with deionized water, precipitated from the aqueous extract using 75% saturated (NH4)2SO4, and subjected on DEAE-cellulose followed by affinity chromatography on sepharose-6B. The activity was tested using hemagglutination assays, and carbohydrate-binding specificity was determined by glycan microarray analysis. Its effects on the mitogenic activity of mouse splenocytes were determined by MTT assay. The novel lectin was adsorbed on ion-exchange chromatography DEAE-cellulose and shown as a band with the molecular mass of 17.5 kDa on a SDS-PAGE and as a single 35.0-kDa peak in gel filtration on Superdex G-75. The hemagglutinating activity of the lectin was inhibited by D-glucose, lactose, D-galactose, and galactosamine. The lectin was stable on 60°C. The hemagglutinating activity of lectin was reduced by 50% at 70°C. At 80°C, it was further reduced to 6.25% of its original activity. The hemagglutinating activity was the highest at pH 6-9. Moreover, its hemagglutinating activity was inhibited by Mg2+ and Ca2+ ions. The lectin isolated from P. ferulae in the current study possessed highly potent hemagglutinating and proliferative activities toward mouse splenocytes.


Subject(s)
Lectins/isolation & purification , Pleurotus/chemistry , Animals , Female , Hemagglutination Tests , Lectins/chemistry , Lectins/pharmacology , Mice , Mice, Inbred BALB C
7.
BMC Musculoskelet Disord ; 13: 34, 2012 Mar 10.
Article in English | MEDLINE | ID: mdl-22405078

ABSTRACT

BACKGROUND: Early diagnosis of osteoarthritis (OA) is essential for preventing further cartilage destruction and decreasing severe complications. The aims of this study are to explore the relationship between OA pathological grades and quantitative acoustic parameters and to provide more objective criteria for ultrasonic microscopic evaluation of the OA cartilage. METHODS: Articular cartilage samples were prepared from rabbit knees and scanned using ultrasound biomicroscopy (UBM). Three quantitative parameters, including the roughness index of the cartilage surface (URI), the reflection coefficients from the cartilage surface (R) and from the cartilage-bone interface (Rbone) were extracted. The osteoarthritis grades of these cartilage samples were qualitatively assessed by histology according to the grading standards of International Osteoarthritis Institute (OARSI). The relationship between these quantitative parameters and the osteoarthritis grades was explored. RESULTS: The results showed that URI increased with the OA grade. URI of the normal cartilage samples was significantly lower than the one of the OA cartilage samples. There was no significant difference in URI between the grade 1 cartilage samples and the grade 2 cartilage samples. The reflection coefficient of the cartilage surface reduced significantly with the development of OA (p < 0.05), while the reflection coefficient of the cartilage-bone interface increased with the increase of grade. CONCLUSION: High frequency ultrasound measurements can reflect the changes in the surface roughness index and the ultrasound reflection coefficients of the cartilage samples with different OA grades. This study may provide useful information for the quantitative ultrasonic diagnosis of early OA.


Subject(s)
Cartilage, Articular/diagnostic imaging , Knee Joint/diagnostic imaging , Microscopy, Acoustic , Osteoarthritis, Knee/diagnostic imaging , Animals , Cartilage, Articular/pathology , Disease Models, Animal , Early Diagnosis , Female , Knee Joint/pathology , Osteoarthritis, Knee/pathology , Predictive Value of Tests , Rabbits , Severity of Illness Index , Surface Properties
8.
Zhonghua Fu Chan Ke Za Zhi ; 46(6): 407-11, 2011 Jun.
Article in Zh | MEDLINE | ID: mdl-21781578

ABSTRACT

OBJECTIVE: To evaluate the efficacy and safety of focused ultrasound ablation in the treatment of submucosal fibroids which broke into uterine cavity less than 50%. METHODS: From Oct. 2006 to Sept. 2009, 66 patients with 69 submucosal fibroids broke into uterine cavity less than 50% diagnosed by MRI in Chinese People's Liberation Army General Hospital were enrolled in this study. They were treated by ultrasound-guided focused ultrasound ablation in the outpatient department, which using the contrast enhanced ultrasonography to assess the efficacy after ablation immediately, to measure reduction of fibroids volume and record adverse effect before and after ultrasound ablation. At 3, 6, 12 and 24 months after treatment, ablation outcome and fibroids volumes were evaluated by contrast ultrasound. The changes of clinical symptom were evaluated by the symptom severity score (SSS) of the uterine fibroid quality-of-life instrument (UFS-QOL). RESULTS: The average volume of fibroids in 66 patients with 68 submucosal fibroids were (151 ± 134) cm(3) before treatment and (114 ± 104) cm(3) no enhanced regional after treatment. The ablation rate of target fibroids was (77 ± 16)%. All patients completed this treatment successfully, they were followed up for 6 - 44 months, the median follow-up time was 24 months. No serious complication was observed. However, there were 52% (34/66) patients presented vaginal discharge after ablation, it disappeared gradually after 3 to 4 menstrual cycles. The SSS and the menstrual period symptom scores were significantly lower than that before ablation at the follow-up of 3, 6, 12 and 24 months, the rates were 20.9%, 38.0%, 45.1%, 47.1% and 42.0%, 63.8%, 64.2%, 68.8%, which all reached statistical difference (P < 0.05). The necrotic fibroids were absorbed gradually, the reduction rates of fibroid volume were 44.7%, 66.0%, 77.7% and 89.8%. CONCLUSION: It was safe and efficacy that focused ultrasound ablation was used in treatment of submucosal fibroids which broke into the uterine less than 50%.


Subject(s)
High-Intensity Focused Ultrasound Ablation/methods , Leiomyoma/therapy , Uterine Neoplasms/therapy , Adult , Female , Follow-Up Studies , Humans , Leiomyoma/diagnostic imaging , Leiomyoma/pathology , Magnetic Resonance Imaging , Middle Aged , Prospective Studies , Treatment Outcome , Ultrasonography/methods , Uterine Neoplasms/diagnostic imaging , Uterine Neoplasms/pathology , Uterus/diagnostic imaging , Uterus/pathology
9.
Neural Regen Res ; 16(1): 143-149, 2021 Jan.
Article in English | MEDLINE | ID: mdl-32788469

ABSTRACT

Veins are easy to obtain, have low immunogenicity, and induce a relatively weak inflammatory response. Therefore, veins have the potential to be used as conduits for nerve regeneration. However, because of the presence of venous valves and the great elasticity of the venous wall, the vein is not conducive to nerve regeneration. In this study, a novel tissue engineered nerve graft was constructed by combining normal dissected nerve microtissue with an autologous vein graft for repairing 10-mm peripheral nerve defects in rats. Compared with rats given the vein graft alone, rats given the tissue engineered nerve graft had an improved sciatic static index, and a higher amplitude and shorter latency of compound muscle action potentials. Furthermore, rats implanted with the microtissue graft had a higher density and thickness of myelinated nerve fibers and reduced gastrocnemius muscle atrophy compared with rats implanted with the vein alone. However, the tissue engineered nerve graft had a lower ability to repair the defect than autogenous nerve transplantation. In summary, although the tissue engineered nerve graft constructed with autologous vein and nerve microtissue is not as effective as autologous nerve transplantation for repairing long-segment sciatic nerve defects, it may nonetheless have therapeutic potential for the clinical repair of long sciatic nerve defects. This study was approved by the Experimental Animal Ethics Committee of Chinese PLA General Hospital (approval No. 2016-x9-07) on September 7, 2016.

10.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 32(4): 470-2, 2010 Aug.
Article in Zh | MEDLINE | ID: mdl-20868613

ABSTRACT

OBJECTIVE: To explore the clinical value of contrast-enhanced ultrasound (CEUS) in the diagnosis of Budd-Chiari syndrome with inferior vena cava (IVC) obstruction. METHOD: A total of 38 patients with Budd-Chiari syndrome with IVC obstruction were examined by CEUS before and after vascular interventional management, and the results were compared with angiographic findings. RESULTS: The location and degree of IVC obstruction were clearly showed on CEUS. Enhancement was found in the occlusion site, and blood flow was narrowed at the stenosis site. The arrival time was earlier after treatment in the IVC obstruction, and it was positively correlated with the pressure of IVC( P< 0.05). CONCLUSION: The location and type of the occlusion and stenosis in IVC can be accurately determined by CEUS. Therefore, CEUS can provide useful information for the selection of surgical procedures and post-operative effectiveness.


Subject(s)
Budd-Chiari Syndrome/diagnostic imaging , Adolescent , Adult , Female , Humans , Male , Middle Aged , Ultrasonography , Vena Cava, Inferior/diagnostic imaging , Young Adult
11.
Zhonghua Yan Ke Za Zhi ; 44(5): 448-54, 2008 May.
Article in Zh | MEDLINE | ID: mdl-18953902

ABSTRACT

OBJECTIVE: To study the development and opsin expression of zebrafish photoreceptors after CRX gene knock-down. METHODS: It was a experimental study. CRX-MOs was microinjected into the zygote of zebrafish. Seventy-two and 96 hours post fertilization, we prepared the eye histological slides, electron microscope samples and whole mount in-situ hybridizations to study the development and opsin expression. of photoreceptors. RESULTS: There were no obvious histological changes in optical microscope observation after CRX-MOs microinjection. Outer segments were developed in control and blank groups, but no obvious outer segment was found in CRX group by electron microscope observation. Opsin Rho, SWS1 and SWS2 were expressed in control and blank groups and the expression of these genes was obviously decreased in CRX group. CONCLUSION: CRX gene can influence the development and opsins expression of photoreceptors outer segments in zebrafish.


Subject(s)
Homeodomain Proteins/genetics , Photoreceptor Cells, Vertebrate , Photoreceptor Cells/cytology , Trans-Activators/genetics , Zebrafish/embryology , Animals , Genes, Homeobox , Zebrafish/genetics
12.
Cardiovasc Res ; 74(2): 290-303, 2007 May 01.
Article in English | MEDLINE | ID: mdl-17367767

ABSTRACT

OBJECTIVE: Cardiac contractility is regulated tightly as an extrinsic and intrinsic homeostatic mechanism to the heart. The molecular basis of the intrinsic system is largely unknown. Here, we test the hypothesis that bone morphogenetic protein-2 (BMP-2) mediates embryonic cardiac contractility upstream of myocyte-specific enhancer factor 2A (MEF2A). METHODS: The BMP-2 and MEF2A expression pattern was analyzed by RT-PCR, Western blotting, whole-mount in situ hybridization, and an in vivo transgenic approach. The cardiac phenotype of BMP-2 and MEF2A knock-down zebrafish embryos was analysed. Cardiac contractions were recorded with a video camera. Myofibrillar organization was observed with transmission electron microscopy. Gene expression profiles were performed by quantitative real-time PCR analysis. RESULTS: We demonstrate that BMP-2 and MEF2A are co-expressed in embryonic and neonatal cardiac myocytes. Furthermore, we provide evidence that BMP-2 is required for cardiac contractility in vitro and in vivo and that MEF2A expression can be activated by BMP-2 signaling in neonatal cardiomyocytes. BMP-2 is involved in the assembly of the cardiac contractile apparatus. Finally, we find that exogenous MEF2A is sufficient to rescue ventricular contractility defects in the absence of BMP-2 function. CONCLUSIONS: In all, these observations indicate that BMP-2 and MEF2A are key components of a pathway that controls the cardiac ventricular contractility and suggest that the BMP2-MEF2A pathway can offer new opportunities for the treatment of heart failure.


Subject(s)
Bone Morphogenetic Proteins/metabolism , MADS Domain Proteins/metabolism , Myocardial Contraction/physiology , Myocytes, Cardiac/physiology , Myogenic Regulatory Factors/metabolism , Signal Transduction/physiology , Transforming Growth Factor beta/metabolism , Animals , Animals, Newborn , Base Sequence , Blotting, Western , Bone Morphogenetic Protein 2 , Bone Morphogenetic Proteins/analysis , Bone Morphogenetic Proteins/genetics , Carrier Proteins/pharmacology , Cells, Cultured , Gene Expression Regulation/drug effects , Genetic Engineering , Heart/embryology , Humans , MADS Domain Proteins/analysis , MADS Domain Proteins/genetics , MEF2 Transcription Factors , Molecular Sequence Data , Myogenic Regulatory Factors/analysis , Myogenic Regulatory Factors/genetics , Organisms, Genetically Modified , RNA Interference , RNA, Small Interfering/administration & dosage , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Transforming Growth Factor beta/analysis , Transforming Growth Factor beta/genetics , Zebrafish/embryology , Zebrafish/metabolism , Zebrafish Proteins/genetics
13.
Chin Med J (Engl) ; 120(13): 1166-71, 2007 Jul 05.
Article in English | MEDLINE | ID: mdl-17637246

ABSTRACT

BACKGROUND: Folic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of folic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase (DHFR) gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 (HAND2) and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations. METHODS: Morpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression. RESULTS: DHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos (P < 0.05). Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos. CONCLUSIONS: DHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.


Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Heart Defects, Congenital/etiology , Heart/embryology , Tetrahydrofolate Dehydrogenase/physiology , Zebrafish Proteins/genetics , Animals , Basic Helix-Loop-Helix Transcription Factors/physiology , Female , Tetrahydrofolate Dehydrogenase/genetics , Zebrafish , Zebrafish Proteins/physiology
14.
Zhongguo Dang Dai Er Ke Za Zhi ; 9(2): 159-63, 2007 Apr.
Article in Zh | MEDLINE | ID: mdl-17448316

ABSTRACT

OBJECTIVE: To study the effect of methotrexate (MTX), a folic acid antagonist which can lead to folic acid deficient, on the cardiac development and on the expressions of BMP2b and HAS2 in zebrafish. METHODS: The zebrafish embryos at 6-48 hrs post fertilization (hpf) were treated with various concentrations of MTX (0.5 x 10(-3), 1.0 x 10(-3) and 2.0 x 10(-3) M). At 48 hpf, the percentage of cardiac malformation and heart rate were recorded. The zebrafish embryos at 6-10 hpf treated with 1.5 x 10(-3) M MTX were used as the MTX treatment group. At 24 and 48 hpf the cardiac morphology was observed under a microscope. The expressions of BMP2b and HAS2 in zebrafish were detected by in situ antisense RNA hybridization and real-time PCR. RESULTS: 6-12 hpf, the early embryonic developmental stage, was a sensitive period that MTX affected cardiac formation of zebrafish. The retardant cardiac development and the evidently abnormal cardiac morphology was found in the MTX treatment group. The results of in situ antisense RNA hybridization showed that the expressions of BMP2b and HAS2 in the zebrafish heart were reduced in the MTX treatment group at 36 and 48 hpf. The real-time PCR results demonstrated that the BMP2b expression decreased at 12, 24, 36 and 48 hpf, and that the HAS2 expression decreased at 24, 36 and 48 hpf in the treatment group compared with the control group without MTX treatment. CONCLUSIONS: The inhibition of folic acid function may affect cardiac development of early embryos, resulting in a retardant development and a morphological abnormality of the heart in zebrafish, possibly by down-regulating the expressions of BMP2b and HAS2.


Subject(s)
Abnormalities, Drug-Induced/etiology , Bone Morphogenetic Proteins/genetics , Folic Acid Antagonists/toxicity , Gene Expression Regulation/drug effects , Glucuronosyltransferase/genetics , Heart Defects, Congenital/chemically induced , Methotrexate/toxicity , Zebrafish Proteins/genetics , Animals , Bone Morphogenetic Protein 2 , Down-Regulation , Hyaluronan Synthases , Polymerase Chain Reaction , Zebrafish
15.
Beijing Da Xue Xue Bao Yi Xue Ban ; 38(1): 62-5, 2006 Feb 18.
Article in English | MEDLINE | ID: mdl-16415969

ABSTRACT

OBJECTIVE: To evaluate the frequency of MSI in epithelial ovarian tumors and its relationship with clinicopathologic features. METHODS: Ninety fresh specimens of epithelial ovarian tumors, including 74 primary and 16 secondary tumors, were collected. Microsatellite analysis was carried out using 5 mono-and dinucleotide markers from the National Cancer Institute Consensus Panel by fluorescence-labeled polymerase chain reaction. RESULTS: Of 90 epithelial ovarian tumors analyzed, 18 demonstrated a high level of microsatellite instability (MSI-H), 30 demonstrated a low level of microsatellite instability (MSI-L), and the remaining 42 exhibited microsatellite stability (MSS). Frequency of microsatellite instability (MSI) at loci BAT-25 was higher than that at any other loci. No correlation was found between MSI level and patient age, tumor type, tumor differentiation (P>0.05). But the microsatellite instability-high phenotype correlates with clinical stage. It tended to occur more frequently in early-stage tumors (P=0.03). CONCLUSION: The frequent MSI in epithelial ovarian tumors suggests that it is an early event to involve in the development of epithelial ovarian tumors.


Subject(s)
Carcinoma/genetics , Microsatellite Instability , Ovarian Neoplasms/genetics , Adult , Female , Humans , Middle Aged
16.
Oncotarget ; 7(14): 17737-50, 2016 Apr 05.
Article in English | MEDLINE | ID: mdl-26894969

ABSTRACT

Tristetraprolin (also known as TTP, TIS11, ZFP36, and Nup475) is a well-characterized tumor suppressor that is down-regulated in several tumor types. In the current study, we found that TTP expression was markedly reduced in pancreatic cancer samples as compared to matched normal tissues. Low TTP level was associated with age (P=0.037), tumor size (P=0.008), tumor differentiation (P=0.004), postoperative T stage (pT stage, P<0.001), postoperative N stage (pN stage, P=0.008) and TNM stage (P<0.001). Moreover, low TTP expression predicted reduced survival rates and poor patient outcome. We also found that TTP impairs pancreatic cancer cell proliferation both in vivo and in vitro. Fluorescence Activated Cell Sorting (FACS) assay showed that TTP over-expression both increases apoptosis and decreases proliferation in pancreatic cancer cells. RNA-sequencing analysis showed that TTP over-expression downregulates several tumor-related factors, including Pim-1 and IL-6. Our findings indicate that TTP could serve as a potential prognostic indicator in pancreatic cancer.


Subject(s)
Pancreatic Neoplasms/metabolism , Tristetraprolin/biosynthesis , Animals , Cell Differentiation/physiology , Cell Line, Tumor , Cell Proliferation/physiology , Heterografts , Humans , Interleukin-6/metabolism , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/pathology , Prognosis , Survival Rate , Transfection , Tristetraprolin/metabolism
17.
FEBS Lett ; 579(21): 4843-50, 2005 Aug 29.
Article in English | MEDLINE | ID: mdl-16107252

ABSTRACT

Myocyte-specific enhancer factor 2A (MEF2A) regulates a broad range of fundamental cellular processes including cell division, differentiation and death. Here, we tested the hypothesis that MEF2A is required in cardiac contractility employing zebrafish as a model organism. MEF2A is highly expressed in heart as well as somites during zebrafish embryogenesis. Knock-down of MEF2A in zebrafish impaires the cardiac contractility and results in sarcomere assembly defects. Dysregulation of cardiac genes in MEF2A morphants suggests that sarcomere assembly disturbances account for the cardiac contractile deficiency. Our studies suggested that MEF2A is essential in cardiac contractility.


Subject(s)
DNA-Binding Proteins/metabolism , Myocardial Contraction/physiology , Myocardium/metabolism , Transcription Factors/metabolism , Zebrafish/embryology , Animals , DNA-Binding Proteins/genetics , Gene Expression , Humans , MADS Domain Proteins , MEF2 Transcription Factors , Microinjections , Morphogenesis , Myocardium/ultrastructure , Myogenic Regulatory Factors , Oligonucleotides, Antisense , Phenotype , Promoter Regions, Genetic , RNA Splicing , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Transcription Factors/genetics , Zebrafish/anatomy & histology , Zebrafish/genetics , Zebrafish/metabolism
18.
Neural Regen Res ; 10(11): 1846-55, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26807123

ABSTRACT

Nerve growth factor (NGF) plays an important role in promoting neuroregeneration after peripheral nerve injury. However, its effects are limited by its short half-life; it is therefore important to identify an effective mode of administration. High-frequency ultrasound (HFU) is increasingly used in the clinic for high-resolution visualization of tissues, and has been proposed as a method for identifying and evaluating peripheral nerve damage after injury. In addition, HFU is widely used for guiding needle placement when administering drugs to a specific site. We hypothesized that HFU guiding would optimize the neuroprotective effects of NGF on sciatic nerve injury in the rabbit. We performed behavioral, ultrasound, electrophysiological, histological, and immunohistochemical evaluation of HFU-guided NGF injections administered immediately after injury, or 14 days later, and compared this mode of administration with intramuscular NGF injections. Across all assessments, HFU-guided NGF injections gave consistently better outcomes than intramuscular NGF injections administered immediately or 14 days after injury, with immediate treatment also yielding better structural and functional results than when the treatment was delayed by 14 days. Our findings indicate that NGF should be administered as early as possible after peripheral nerve injury, and highlight the striking neuroprotective effects of HFU-guided NGF injections on peripheral nerve injury compared with intramuscular administration.

19.
Beijing Da Xue Xue Bao Yi Xue Ban ; 36(5): 519-24, 2004 Oct.
Article in Zh | MEDLINE | ID: mdl-15489935

ABSTRACT

OBJECTIVE: To analysis the relationship between glucocorticosteroids (GCS) usage and side effects in the treatment of severe acute respiratory syndrome (SARS). METHODS: All clinical records of probable SARS patients in Beijing were collected and input into an Epi6 database, in which 1 291 patients had entire information and met the clinical criteria of SARS. The usage of GCS and GCS associated side effects were analyzed retrospectively. RESULTS: Patients accepted GCS therapy were 83.96% (n=1 084), whereas 16.04%(n=207) did not take GCS. The average dosage of GCS was 160 mg/d in the first week, and then reduced to 80 mg/d and 40 mg/d in the second and the third weeks, respectively. Initial blood glucose, systolic pressure (SBP), and diastolic pressure (DBP) were no significant difference between GCS group and non-GCS group. The highest blood glucose during the treatment in GCS group was markedly higher than that in non-GCS group [(8.68+/-4.80) mmol/L vs (6.39+/-3.71) mmol/L, P<0.05)]. The highest blood glucose and average blood glucose after initiation of GCS therapy were elevated in GCS group. The levels of blood glucose were correlated with the initial, maximum, and cumulative GCS dosages. Average blood glucose was increased markedly in groups with MP(Initial) > or =80 mg/d (Methyprednisonlone), MP(Maximal) > or =160 mg/d, or MP(Cumulative) > or = 3000 mg. The blood glucose grew up significantly in the first and the second weeks in the treatment with GCS, and then returned to normal level gradually. Hyperglycemia duration in the group with MP(Cumulative) > or =3000 mg persisted longer than that in the other groups (P< 0.05). The lowest serum potassium during the treatment and the duration of hypokalemia in GCS group were significantly different from that in non-GCS group [(3.66+/-0.50) mmol/L vs (4.01+/-0.51) mmol/L, P< 0.001 ;1(1, 75) days vs 1(1, 9) days, P<0.05, respectively]. Average serum potassium and the duration of hypokalemia were related to the dosages of GCS. Serum potassium reached its nadir in the first week of GCS treatment and then grew up in the second week. In groups with MP(Initial) > or =320 mg/d, MP(Maximal) > or =320 mg/d, and MP(Cumulative) > or =3000 mg, the level of serum potassium was lower and the duration of hypokalemia was longer than that in other groups. They began to returned to normal level in the third week. Administration of GCS prolonged the time of hypocalcemia[19 (1, 74) days in GCS group vs 8 (1, 32) days in non-GCS group, P< 0.05]. The duration of hypocalcemia was prolonged according to the increasing of the maximal or the cumulative dosage of GCS. However, the duration of hypocalcemia in group with MP(Cumulative) <999 mg was similar to that in non-GCS group (P > 0.05). After GCS administration, SBP and DBP were increased gradually, and reached their peaks in the fourth week [SBP(117.2+/-14.0) mm Hg and DBP (72.5+/-9.1) mm Hg vs SBP (120.0+/-12.5) mm Hg and DBP (74.5+/-8.7) mm Hg, P< 0.05, 1 mm Hg=0.133 kPa]. CONCLUSION: Hyperglycemia and hypokalemia are correlated with GCS dosage and duration. Administration with GCS influences SBP, DBP, and duration of hypocalcemia. Appropriate low dosage of GCS (MP(Initial) and MP(Maximal) < 159 mg/d, MP(Cumulative)< 2999 mg) causes few changes of blood glucose, serum potassium, and blood calcium. It is important to monitor laboratory findings during the treatment with GCS.


Subject(s)
Glucocorticoids/adverse effects , Hyperglycemia/chemically induced , Hypokalemia/chemically induced , Severe Acute Respiratory Syndrome/drug therapy , Dose-Response Relationship, Drug , Female , Glucocorticoids/therapeutic use , Humans , Hypocalcemia/chemically induced , Male , Methylprednisolone/adverse effects , Methylprednisolone/therapeutic use , Retrospective Studies
20.
Zhongguo Zhen Jiu ; 33(11): 1028-32, 2013 Nov.
Article in Zh | MEDLINE | ID: mdl-24494297

ABSTRACT

UNLABELLED: OBJECTIVEL to study the visualization of acupotomy therapy and to summarize differences of interventional ultrasound combined with acupotomy on spinal and articular diseases. METHODS: With randomized block design, 20 cases of shoulder periarthritis, 20 cases of knee arthritis, 20 cases of lumbar disc herniation and 20 cases of cervical disc herniation were divided into an ultrasound-guided group and regular group, 10 cases of each diseas in each group. The ultrasound-guided acupotomy and regular acupotomy were given for one time, respectively. Efficacy was observed after one week. Visual analogue scale (VAS) and fuction score were applied for efficacy assessment, including Wang's funtion evaluation table for cervical disc herniation, modified lumbago assessment by Japanese Orthopaedic Association for lumbar disc herniation, modified lumbago assessment arthritis and CMC shoulder funtion scale for shoulder periarthritis. The changes of scores of patients in two groups were observed before and after the treatment. RESULTS: Each disease in each group achieved favorable efficacy; compared before and after treatment, differences of VAS and each function score all were statistically significant (allP<0.01). The efficacy of lumbar disc herniation and cerical disc herniation in two groups was not obviously different (both P>0.01). The efficacy of lumbar disc herniation and cervical disc herniation in two groups was not obviously different (bothP>0.05), while VAS and fuction scores improvements of shoulder periarthritis and knee +/-5.75 vs 75.72+/-8.56; knee arthritis: 90.40+/-7.35 vs 75.54+/-9.21, both P<0.05). CONCLUSION: Whether ultrasound guidance is applied during acupotomy therapy made no obvious influence on efficacy of lumbar disc herniation and cervical disc herniation, but for shoulder periarthritis and knee arthritis better results could be acchieved with ultrasound guidance.


Subject(s)
Acupuncture Therapy , Spinal Diseases/therapy , Acupuncture Points , Acupuncture Therapy/instrumentation , Adult , Aged , Female , Humans , Male , Middle Aged , Spinal Diseases/diagnostic imaging , Treatment Outcome , Ultrasonography
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