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As a pattern recognition receptor, CD36 antigen participates in a series of pathophysiological processes, and has well been documented in transfusion medicine. This article reviews the discovery, structure and expression of CD36, the type and frequency of CD36 antigen deletion, as well as the relationship between anti-CD36 and transfusion-related immune diseases.
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BACKGROUND:Bone transplantation is the only method for the repair of bone defects. However, traditional bone transplantation has some disadvantages. Bone tissue engineering, as a new treatment strategy, can achieve the desire therapeutic outcomes. OBJECTIVE:To fabricate a new tissue-engineered scaffold for improving bone repair effectively. METHODS:Hydroxyapatites (HA) with different Ca/P (1.50/1.67) ratios were synthesized by chemical precipitation method and microwave radiation method. Composite scaffolds of palygorskite (APC)/HA/polycaprolactone (PCL)/col agen (COL), APC/calcium deficiency HA (CDHA)/PCL/COL, and APC/PCL/COL (control group) were prepared by solution perfusion-solvent evaporation and ion leaching method. The material characterization, active ingredients, hydrophilic property, and mechanical properties were evaluated by scanning electron microscope, infrared spectrometer, surface contact measuring instrument and universal mechanics, respectively. The histocompatibility of the implant with the host was assessed through animal experiments. RESULTS AND CONCLUSION:By precise control of pH range, HA with different Ca/P ratios could be synthesized. The mechanical properties, air permeability, hydrophilic property of the APC/HA/PCL/COL and APC/CDHA/PCL/COL composite materials were significantly increased compared with the APC/PCL/COL composite material (P<0.05), while the porosity, water absorption expansion rate were significantly decreased (P<0.05). Results from our animal experiments showed that no immune inflammatory reaction was observed suggesting that the composite materials hold good histocompatibility. To conclude, the APC/HA (1.50/1.67)/PCL/COL composite materials are promising bone substitutes in bone tissue repair.
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Objective To investigate the features and treatments of teardrop fracture of the axis.Methods Of the 17 consecutive patients with teardrop fracture of the axis who had been managed between January 2008 and January 2016 at our trauma center,13 were included in this study according to our research criteria.On their lateral X-ray films of the skull base,the height,width,lateral displacement and rotation of the fracture fragments were measured.Continuity of the anterior longitudinal ligament and instability of C2-3 and posterior ligamentous complex were evaluated on their cervical MRI images.Seven patients were immobilized for 3 months with the Philadelphia collar or Halo-vest device and 6 ones underwent anterior C2-3 cervical surgery.Results For patients receiving conservative and operative treatments,at the sagittal view,the height,width,lateral displacement,anterior rotation and posterior displacement of the fracture fragments averaged 12.0 mm versus 14.8 mm,6.85 mm versus 8.33 mm,7.07 mm versus 8.50 mm,20.0° versus 30.1°,and 1.71 mm versus 3.0 mm,respectively.One patient suffered C2 disc injury and 6 ones C3 disc injury.All the patients were followed up for an average of 26.4 months (from 12 to 36 months).Complications included uncomfortable swallowing in 3 cases and mild residual neck pain in one.There was no delayed union,nonunion,or vertebral instability.At last follow-ups,the mean visual analogue score for pain was 1.7 and the Japanese Orthopaedic Association scores were 17 in 11 patients and 16 in 2 patients.Conclusions Most teardrop fractures can be treated conservatively because their small fracture fragments and minor displacements can be reduced after traction.However,those with large fragments and C2-3 vertebral injury and instability should be treated by anterior cervical discectomy and fusion.
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Objective To investigate the features and treatments of teardrop fracture of the axis.Methods Of the 17 consecutive patients with teardrop fracture of the axis who had been managed between January 2008 and January 2016 at our trauma center,13 were included in this study according to our research criteria.On their lateral X-ray films of the skull base,the height,width,lateral displacement and rotation of the fracture fragments were measured.Continuity of the anterior longitudinal ligament and instability of C2-3 and posterior ligamentous complex were evaluated on their cervical MRI images.Seven patients were immobilized for 3 months with the Philadelphia collar or Halo-vest device and 6 ones underwent anterior C2-3 cervical surgery.Results For patients receiving conservative and operative treatments,at the sagittal view,the height,width,lateral displacement,anterior rotation and posterior displacement of the fracture fragments averaged 12.0 mm versus 14.8 mm,6.85 mm versus 8.33 mm,7.07 mm versus 8.50 mm,20.0° versus 30.1°,and 1.71 mm versus 3.0 mm,respectively.One patient suffered C2 disc injury and 6 ones C3 disc injury.All the patients were followed up for an average of 26.4 months (from 12 to 36 months).Complications included uncomfortable swallowing in 3 cases and mild residual neck pain in one.There was no delayed union,nonunion,or vertebral instability.At last follow-ups,the mean visual analogue score for pain was 1.7 and the Japanese Orthopaedic Association scores were 17 in 11 patients and 16 in 2 patients.Conclusions Most teardrop fractures can be treated conservatively because their small fracture fragments and minor displacements can be reduced after traction.However,those with large fragments and C2-3 vertebral injury and instability should be treated by anterior cervical discectomy and fusion.
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The present study developed an oral hepatocyte growth factor (HGF) gene therapy strategy for gastric ulcers treatment. An attenuated Salmonella typhimurium that stably expressed high HGF (named as TPH) was constructed, and the antiulcerogenic effect of TPH was evaluated in a rat model of gastric ulcers that created by acetic acid subserosal injection. From day 5 after injection, TPH (1 × 10⁹ cfu), vehicle (TP, 1 × 10⁹ cfu), or sodium bicarbonate (model control) was administered orally every alternate day for three times. Then ulcer size was measured at day 21 after ulcer induction. The ulcer area in TPH-treated group was 10.56 ± 3.30 mm², which was smaller when compared with those in the TP-treated and model control groups (43.47 ± 4.18 and 56.25 ± 6.38 mm², respectively). A higher level of reepithelialization was found in TPH-treated group and the crawling length of gastric epithelial cells was significantly longer than in the other two groups (P < 0.05). The microvessel density in the ulcer granulation tissues of the TPH-treated rats was 39.9 vessels/mm², which was greater than in the TP-treated and model control rats, with a significant statistical difference. These results suggest that TPH treatment significantly accelerates the healing of gastric ulcers via stimulating proliferation of gastric epithelial cells and enhancing angiogenesis on gastric ulcer site.
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Animals , Rats , Acetic Acid , Epithelial Cells , Genetic Therapy , Granulation Tissue , Hepatocyte Growth Factor , Hepatocytes , Microvessels , Models, Animal , Salmonella typhimurium , Salmonella , Sodium Bicarbonate , Stomach Ulcer , UlcerABSTRACT
BACKGROUND:Although there is a certain progress in the preparation of tissue-engineered bone tissue using a variety of materials, some deficiencies have appeared such as mismatching between scaffold degradation rate and new bone formation rate, slow tissue growth, toxic metabolites. OBJECTIVE:To build a new type of inducible bone repair composite scaffold with bionic bone structurematerials and to evaluate its physicochemical and biological properties. METHODS: Icarin encapsulated by chitosan was used to prepare drug-loaded microspheres, and the drug release rate of the microspheres was detected. Chitosan microspheres were mixed with colagen to build the core part of scaffold materials. Hydroxyapatite (HA), polycaprolactone (PCL) and colagen were mixed in hexafluoride isopropanol (HFIP) to prepare the HA/PCL/colagen outer part of composite scaffold material at the rate of 0:3:3, 1:3:3, 2:3:3, 3:3:3. Each proportional electrospinning was used for one layer, and finaly the 4-layer outer tube of the scaffold was produced. The tube core and outer tube were crosslinked by 1% genipin. Universal material testing machine, surface contact angle meter, infrared spectroscopy, scanning electron microscope, water absorption, permeability, porosity,in vitro degradation tests for cross-linked and uncross-linked were used to observe the structure and characteristics of tubular materials. Bone marrow mesenchymal stem cels were seeded on the surface of cross-linked and uncross-linked bone repair materials to evaluate the biocompatibility of the scaffolds. Cross-linked and uncross-linked bone repair materials were implanted subcutaneously into Wistar rats to evaluate the histocompatibility of the scaffolds. RESULTS AND CONCLUSION:The drug in the scaffold had a suitable release; the bone scaffold material had good uniformity, and cross-linked scaffolds materials had better mechanical properties, water absorption and permeability than the uncross-linked(P < 0.05). The degradation rate of the cross-linked group was significantly lower than that of the uncross-linked group (P< 0.05). Hematoxylin-eosin staining showed that the bone marrow mesenchymal stem cels could adhere wel to the cross-linked and uncross-linked materials. No inflammatory reactions occurred after subcutaneous implantation of cross-linked and uncross-linked materials. These findings indicate that the cross-linked scaffold for inducible bone tissue engineering has good biocompatibility and mechanical properties.
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BACKGROUND:Although the preparation of bone tissue engineering scaffolds can achieve satisfactory results by solvent casting/particulate leaching, in situ molding method, electrospinning, phase seperation/freeze drying, gas foaming, there are stil some deficiencies in the accuracy, pore uniformity, spatial structure complexity, personalized stents. ＜br> OBJECTIVE:To prepareβ-tricalcium phosphate bone tissue engineering scaffolds using 3D printing. ＜br> METHODS:Drug-loadedβ-tricalcium phosphate scaffolds were prepared with 3D printing, and the structure was observed to measure its porosity and mechanical strength. The scaffold was immersed in simulated body fluid for 15 weeks to observe the quality change. The scaffold was co-cultured with rat bone marrow mesenchymal stem cells for 7 days to observe celladhesion and morphological changes. Rat bone marrow mesenchymal stem cells were cultured in extracts of drug-loadedβ-tricalcium phosphate scaffold and low-glucose Dulbecco's modified Eagle’s medium containing 15%fetal bovine serum for 24, 48, and 72 hours, to determine the absorbance values and cytotoxicity grading, respectively. Meanwhile, the cells were subjected to osteogenic culture for 1 week, and ＜br> the alkaline phosphatase activities in two groups were detected. ＜br> RESULTS AND CONCLUSION:The prepared scaffold showed irregular micropores, high porosity, uniform pore distribution, high pore connectivity rate, and large compressive strength. The drug-loadedβ-tricalcium phosphate scaffold degraded completely with 15 weeks, and cancellous bone defect repair was completed in the same period. Rat bone marrow mesenchymal stem cells adhered to the surface of drug-loadedβ-tricalcium phosphate scaffold and went deep into the scaffold, showing good growth and proliferation. The activity of alkaline phosphatase was also improved. These findings indicate that the drug-loadedβ-tricalcium phosphate scaffold has good biocompatibility.
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BACKGROUND:Our previous studies have shown that salidroside can induce bone marrow mesenchymal stem cells directly into neuron-like cells, and Ca2+signal is one important way to achieve its biological signal transduction. OBJECTIVE:To investigate the role and mechanism of the calcium/calmodulin (Ca 2+/CaM) signaling pathway inducing bone marrow mesenchymal stem cells to differentiate into nerve cells. METHODS:Bone marrow mesenchymal stem cells were divided into two groups:control groups and salidroside groups. Salidroside groups were induced with different concentrations of salidroside (5, 10, 20, 50 and 100 mg/L) for 24 hours and 100 mg/L salidroside was added to culture cells for different time (12, 24, 48 and 72 hours). Western blot assay was used to detect the expression levels of neural cellmarker, microtubule-associated protein 2, and the important protein of Ca2+/CaM signaling pathway:CaM and calmodulin dependent kinase II (CaMK II). Then Ca2+/CaM signaling pathway specific blockers were applied to cells respectively for 30 minutes, including 500 μmol/L EGTA (Ca 2+chelator), 1 mmol/L Nifedipine(L-type Ca2+channel blocker) and 10 mmol/L LY294002 (PI3K inhibitor). Then, 100 mg/L salidroside was added and cultured for 24 hours. Western blot assay was used to detect the expression of neuron-specific enolase and CaM in the Ca2+/CaM signaling pathway. RESULTS AND CONCLUSION:(1) After inducing with salidroside, the expression of microtubule-associated protein 2 were upregulated (P<0.01), indicating that salidrosid can induce the neuronal differentiation of bone marrow mesenchymal stem cells. (2) After different concentrations of salidrosid induced bone marrow mesenchymal stem cells for 24 hours, the expressions of CaM and CaMK II were significantly upregulated in the 10 mg/L group ( P<0.01);For the 100 mg/L salidrosid that was added for cellinduction for different time, the expressions of CaM and CaMK II were significantly downregulated in 72-hour group (P<0.01). (3) After blocking extracellular Ca2+and PI3K signaling pathway, the expressions of neuron-specific enolase and CaM were higher than those in salidrosid groups (P<0.05). These results suggest that salidrosid can induce bone marrow mesenchymal stem cellto directly differentiate into nerve cells by inhibiting the Ca2+/CaM signaling pathway.
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The objective of this investigation was to study the characteristics and biocompatibility of collagen/polyvinyl alcohol (PVA) membrane crosslinked by UV-riboflavin. Membranes that were made into complex ones with different mass ratios of collagen to PVA (1:1 and 2:1) were synthesized, and crosslinked with UV-riboflavin. The surface characteristics were analyzed using the omnipotent materials instrument, IR, SEM, water absorption test, gas permeability test, and degradation test, respectively. The biocompatibility of membrane complex and rat bone marrow mesenchymal stem cells (BMSCs) was evaluated after 7 d and 14 d, respectively. The collagen/PVA complex membranes showed good homogeneity, mechanical property, degradation ratio, water absorption, gas permeability, etc. The biocompatibility of the collagen/PVA (2:1) complex membrane crosslinked with UV-Riboflavin was higher than that of without crosslinking and collagen/PVA (1:1) membrane. It could be well concluded that collagen/PVA complex membranes crosslinked with UV-riboflavin would have a potential application in biomedicine.
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Animals , Rats , Biocompatible Materials , Chemistry , Collagen , Chemistry , Cross-Linking Reagents , Chemistry , Materials Testing , Membranes, Artificial , Mesenchymal Stem Cells , Cell Biology , Polyvinyl Alcohol , Chemistry , Riboflavin , Chemistry , Tissue Engineering , Methods , Ultraviolet RaysABSTRACT
To investigate the role of the extracellular signal-regulated kinase (ERK1/2) and PI3K/AKT/ mTOR signal pathway inducing bone marrow mesenchymal stem cells (BMSCs) differentiation into neural cells, mouse bone marrow-derived mesenchymal stem cell lines D1 cells were used as research object. And they were divided into control groups and salidroside (SD) groups. Different concentrations (5, 25, 50, 100 and 200 microg x mL(-1) of SD were used and SD (100 microg x mL(-1)) was used to induce at different time (0.5, 1, 3, 6, 9, 12, 24, 48 and 72 h). The immunofluorescence staining chemical technology, real-time PCR and Western blotting were used to detect the positive rates of NSE, MAP2, beta-Tubulin III, NES, GFAP and the expression levels of beta-Tubulin III, NSE, ERK1/2, AKT. The expression of ERK1/2 and NSE was detected when the ERK1/2 and PI3K/AKT/ mTOR signal pathway was blocked by PD98059 and LY294002. It indicated that the positive rates of NSE, MAP2, beta-Tubulin III, NES and GFAP were gradually enhanced with time increased. The expression level of NSE and beta-Tubulin III protein were significantly higher than those in control groups (P < 0.01). The expression of ERK1/2, AKT mRNA and protein were higher with concentration and time increased. When the ERK1/2 and PI3K/AKT/mTOR signal pathway were blocked, the expression levels of NSE, NES and beta-Tubulin III mRNA and NSE protein were inhibited significantly. It points out that SD can stimulate the ERK1/2 and PI3K/AKT/mTOR signal pathway to promote BMSCs differentiation into neural cells.
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The objective of this research was to fabricate a novel tissue inducible nerve guide conduit, and to evaluate its biologic property. The microspheres were prepared with chitosan that encapsulated ligustrazine. The drug release of the chitosan microspheres was detected with application of the controlled release method in vitro. Chitosan microspheres were mixed with collagen to fabricate the tissue inducible nerve conduit, which were crosslinked with 2% genipin for 24h. Mechanical properties of the nerve guide conduit samples, including maximum load and breaking load, were measured using an Instron Series IX Automated Materials Testing System. The flexibility of the nerve guide conduit was determined with the texture evaluation instrument. Different methods, such as scanning electron microscope (SEM), light microscope (LMS) and immunofluorescence were used to analyze the spatial structure of the nerve guide conduit, the distribution of the microspheres, the state of the nerve duct combined with mesenchymal stem cells (MSCs), and the effect of the ligustrazine that released from chitosan microsphere on MSCs differentiation into nerve cells, respectively. The results showed that the chitosan microspheres had better releasing effect. The mechanical properties resultant nerve guide conduit were determined. The maximum load and breaking load of the genipin crosslinked samples were significantly higher than that observed with the non-crosslinkers, increasing to (0.76 +/- 0.15) N and (0.69 +/- 0.17) N from (0.23 +/- 0.09) N and (0.20 +/- 0.12) N for the non-crosslinkers (P < 0.01). The degradation rates of non-crosslinked and crosslinked by genipin were(58.62 +/- 7.59) mg and (9.23 +/- 2.47) mg, respec- tively. This had a statistical significance (P < 0.01). The average linearities in dry and hygrometric state of the nerve guide conduit were (0.597 +/- 0.012) LC and (0.333 +/- 0.015) LC, respectively, which also had statistical significance (P < 0.01). The flexibility in the hygrometric state of the nerve guide conduit was better than that of the dry. SEM analysis of the samples demonstrated that the structures of the nerve guide conduit were significantly changed in crosslinking samples, the microspheres were uniformly distributed on the surface of scaffold, the ligustrazine that released from the chitosan microspheres could promote MSCs to express NSE and MAP2 that were the relevant marker molecule of nerve cells. The nerve guide conduit is combined with MSCs, which promote MSCs proliferation and NSE expression by the ligustrazine that released from the chitosan microspheres. The conduit has better biological compatibility and tissue inducible function.
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Animals , Rats , Cell Differentiation , Cells, Cultured , Chitosan , Chemistry , Collagen , Chemistry , Guided Tissue Regeneration , Methods , Mesenchymal Stem Cells , Cell Biology , Microspheres , Nerve Regeneration , Pyrazines , Pharmacology , Tissue Engineering , Tissue Scaffolds , ChemistryABSTRACT
Alzheimer disease (AD) is a chronic neurodegenerative disease that is characterized by its gradual progression. At present, the cause and mechanism of AD are yet unclear, and there is no effective therapy for treating it. With development of global aging, the prevalence rate of AD is increasing. The life quality of elderly people is affected severely by AD that is ultimately life-threatening. Recently, study on treating AD with traditional Chinese medicine (TCM) has deepened.
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Ammopiptanthus mongolicus shows very strong resistance to severe environments. To isolate drought-resistant genes and elucidate drought-resistant molecular mechanisms of the plant, we constructed a drought-induced full-length cDNA library using SMART (Switching mechanism at 5'-end of RNA transcript) technique. The phage titer of the unamplified library was 1.6 x 10(7) PFU/mL; the recombination percentage was 97.7%; and the sizes of most cloned cDNA fragments were around 1 kb. Three thousand positive clones were randomly selected and sequenced from their 5' ends, and a total of 1 450 Unigenes were identified. By Blast searches against the Nt, Nr and Swissprot databases, we found that 919 Unigenes (amount to 63.4%) showed significant similarity to the annotated genes, and the remaining 531 Unigenes (amount to 36.6%) represented novel genes without any annotation. Among the functional categories of the GO (Gene Ontology) classification, the terms related to physiological process, cellular process, binding, catalytic activity and cellular components were dominant. The next abundant terms were for organelle, protein complex, transporter activity and structural molecule activity. In addition, there were a significant proportion of the terms involved in stimulus response, gene expression regulation, regulation of physiological and biochemical processes and signal transduction. Many of the annotated Unigenes were found to be related to plant resistance to abiotic stresses, and expression analyses of 6 out of these genes by semi-quantitive RT-PCR confirmed their involvements in the response of A. mongolicus to drought stress. These results laid a foundation for the expression profile analysis and the cloning and characterization of drought-resistant genes from the plant in the future.
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Droughts , Fabaceae , Genetics , Gene Expression Regulation, Plant , Gene Library , Genotype , Sequence Analysis, DNA , Stress, Physiological , GeneticsABSTRACT
The present research was aimed to investigate the effects of sinusoidal electromagnetic fields (SEMFs) on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells in rats (rBMSCs) and to find out the intensity with the best therapeutic efficacy. Primary rat bone marrow mesenchymal stem cells were obtained from Wistar rats and screened by the adhesive method. The rBMSCs were exposed to sinusoidal electromagnetic fields with 50Hz frequency and intensities of 0 mT, 1.4 mT, 1.6 mT, 1.8 mT, 2.0 mT, and 2.2 mT respectively, 30 min per day. The proliferation of the rBMSCs was analyzed by MTT reduction assay. The osteogenic differentiation markers including ALP activity, calcium deposition, mineralized bone modulus and collagen I expression were compared between the rats in the exposed groups and those in the control group. The total cellular RNA was extracted after 6, 12, 24 and 48 hours, respectively. The gene expression of Osterix and IGF-1 was examined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The absorbance of exposed groups was suppressed significantly in comparison with that in the control group. The exposure to the rBMSCs with intensity of 1.8 mT strongly enhances the osteogenic differentiation of rBMSCs, indicated by remarkably improved ALP activity, calcium deposition, collagen I expression and the number of mineralized bone nodules compared to that in the control group and other groups. Osterix and IGF-1 were also significantly improved (P < 0.05). The SEMFs with frequency and 50Hz and 1.4-2.2 mT intensities enhanced the osteogenic differentiation of rBMSCs, but inhibited their proliferation in the presence of 0.1% serum culture. Among the rBMSCs used in the tests, the one with 1.8 mT had the strongest activity, indicating that it could be the optimal intensity for the clinical application.
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Animals , Rats , Alkaline Phosphatase , Metabolism , Bone Marrow Cells , Cell Biology , Radiation Effects , Cell Differentiation , Radiation Effects , Cell Proliferation , Radiation Effects , Cells, Cultured , Electromagnetic Fields , Mesenchymal Stem Cells , Cell Biology , Radiation Effects , Osteoblasts , Cell Biology , Radiation Effects , Rats, WistarABSTRACT
Objective To study effects of salmon calcitonin and Heng Gu bone healing reagent on vertebral osteoporotic fracture(OPF).Methods From Nov.2007 to Dec.2009,82 cases of vertebral OPF were treated.These cases were randomly divided into treatment group and control group.The treatment group(42 cascs)were treated with salmon calcitonin and Heng Gu bone healing reagent.The control group(40 cases)received salmon calcitonin only.Pain relief of the 2 groups wag compared.Results Before treatment,the 82 patients were scored 6-9 points by visual analogue scales(VAS)and pain scores of the 2 groups were similar (P>0.05).3 days,5 days,8 days and 15 days after treatment,VAS scores of the 2 groups were significantly different(P<0.01).Compared to the control group,the treatment group had pain relief in shorter time and their bone mineral density Was better showed by the 3-month-later review.Conclusion The combination therapy of salmon calcitonin and Heng Gu bone healing reagent in treating vertebral OPF has better effect of pain relief and bone formation,which is a safe and effective method.
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BACKGROUND: Vertebrae axial rotation is a basic deformity of scoliosis, the rotational degree of which is hard to measure due to the field limitation of posterior spinal instrumentation. Currently, vertebrae rotational degree is measured according to preoperative X-ray film or CT, while no reports concerning measuring vertebrae rotational degree during operation. OBJECTIVE: To explore the feasibility of measurement of vertebrae rotational degree with the entry point of pedicle screws.METHODS: Design of the path of pedicle screws on CT scans before surgery, a line bisection and perpendicular to another connecting bilateral entry point of pedicle screws, and the angle of vertebral rotation (EPPsag) was taken as the angle between this line and the saggital plane. The difference among vertebrae rotational degrees measured by conimeter, Ho's method and EPPsag was compared by Wilcoxon signed rank test. The intra-observer and inter-observer difference was analyzed with One-WayANOVA. Conimeter was used to measure vertebrae rotational degree of each vertebra in 9 lumbar specimens, and the results was compared to EPPsag.RESULTS AND CONCLUSION: There was no significant difference among EPPsag, actual rotational degree and measuring results of Ho's method (P>0.05). The One-Way ANOVA showed that the differences between intra-observer analysis and inter-observer analysis (P>0.05). The results demonstrated that EPPsag can exhibit vertebrae rotational degree accurately and repeatability. This anger can be obtained accurately with the instrument if the vertebrae rotational degree not exceeding 30°.
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BACKGROUND:Tranditional Chinese medicine,which possesses anti-oxidation properties,can promote directional differentiation of mouse bone marrow mesenchymal stem cells(BMMSCs)into nerve cells.Salidrosides,as the effective constituent of Rhodiola,have strong anti-oxidation function.OBJECTIVE:To investigate the molecule mechanism of salidrosides induced differentiation of mouse BMMSCs into nerve cells.METHODS:When in vitro cultured BMMSCs reached 80% confluency,the cells were assigned into 3 groups.Cells in the control group were cultured by complete culture medium;those in the induction and positive control groups were cultured by complete culture medium adding 20 mg/L salidrosides or 0.1 mg/L nerve growth factors(NGF).The related gene and protein of nerve cells were detected using RT-PCR and Western blot method at 12 hours after culture.After that,the cells in the induction group were divided into 3 groups,the blocking agents EGTA(Ca~(2+) chelator),Nifedipine(L-type Ca~(2+) channel blocker)and LY294002(IP3 receptor blocking pharmacon)were applied to block the cellular Ca2* signal pathway respectively for 12 hours.RT-PCR and Western blot methods were used to study the signal transduction of the salidrosides.RESULTS AND CONCLUSION:①The expression of neuron specific enolase(NSE),B-Tubulin III,Nurri mRNA could be found ir the induction and positive control groups,instead of the control group;The expression abundance of the positive control group was smaller than that of the induction group.The expression abundance of GFAP mRNA was very low in each group,but the c-fos mRNA was expressed abundantly in the induction group.②Compared with the positive control group,the induction group could promote the NSE expression obviously,which was no expressed in the blank control group.?The expression of NSE and Nurri were conspicuously down-regulated when the Ecto Ca~(2+) and L-type Ca~(2+) channel and IP3 receptor were blocked respectively.Salidrosides can induce the differentiation BMMSCs into nerve cells. Ca~(2+) signaling and IP3 dependent Ca~(2+) signaling pathway play an important role in transduction salidrosides signal in BMMSCs differentiation.
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<p><b>OBJECTIVE</b>To evaluate the histological changes on the femoral heads of the SANFH rabbit animal models and after it were intervened by Osteoking (herbs of the Yi minority in Yunnan province) using general and light microscope observation.</p><p><b>METHOD</b>A total of 150 New Zealand white rabbits were randomly divided into a non-treatment control group (A group, n = 24), normal rabbits with Osteoking treatment group (B group, n = 24), and the experimental group (n = 102). The experimental group was injected with escherichia coli endotoxin (10 microg x kg(-1)) into auricular vein twice by 24-hour intervals, and prednisolone (20 mg x kg(-1)) was injected into buttock three times by 24-hour intervals to make steroid-induced femoral head necrosis model. At the fifth week, 48 out of 53 rabbits were equally divided into model group (C group, n = 24, models with non-treatment Osteoking) and abnormal rabbits with Osteoking treatment group (D group, n = 24). B group and D group were intragastrically administrated with Osteoking, once every two days. A group and C group were intragastrically administrated with the equal volume of saline. At 8th, 12th and 16th week after model preparation, the femoral head specimens were observed under the general and a light microscope.</p><p><b>RESULT</b>Macroscopic and light microscopic analysis showed that, clear bone necrosis of femoral head was observed in the C group, and a large number of fat cell proliferation was found in the bone marrow cavity. As compared with C group, the damage level of cells in D group was milder, however, the density of bone trabecula from Osteoking treatment was high, and the ratio of bone lacuna was very low. It is also demonstrated that the surface area of bone necrosis was decreased, and the number of cells from adiposities was reduced significantly. The phenomenon of bone necrosis repaired apparently. The morphology of femoral head from A group and B group is normal.</p><p><b>CONCLUSION</b>It suggested that Osteoking could effectively help repair steroid-induced femoral head necrosis in the early stage.</p>
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Animals , Female , Humans , Male , Rabbits , Bone Density , Disease Models, Animal , Drugs, Chinese Herbal , Femur Head , Pathology , Femur Head Necrosis , Drug Therapy , Pathology , Microscopy , Random AllocationABSTRACT
BACKGROUND: Basic fibroblast growth factor (bFGF) has significant promotion effects on repair in trauma, but local application cannot play a role for a long time.OBJECTIVE: To observe expression of bFGF gene in rabbit bone marrow mesenchymai stem cells (BMSCs) following transfection.DESIGN, TIME AND SETTING: The cytogene in vitro observation was performed at the College of Life Science, Yunnan University from March to August 2009.MATERIALS: One Japan flap-eared rabbit was purchased from the Department of Animal, Kunming Medical College. pCDNA3.1plasmid (Invitrogen, USA) was used in this study.METHODS: Bone marrow was extracted from rabbit anterior superior iliac spine. BMSCs were harvested by the adherent method.Cells were digested and subcultured when 80% confluent. According to GeneBank bFGF cDNA sequence, gene was designed and synthesized. Following electropherosis, the gel was retrieved using xhol I, BamH I enzyme digestion. Restriction enzyme was used to perform enzyme digestion, electropherosis and gene recovery in PcDNA Vector plasmid. bFGF DNA was connected with PcDNA Vector plasmid. PcDNA-bFGF eukaryotic expression vector was constructed. Recombinant was transfected into rabbit BMSCs using liposome infection protocol, and stable transfected line was screened.MAIN OUTCOME MEASURES: BMSC surface antigen expression was measured. Western blot was utilized to determine the expression of target protein.RESULTS: Results of flow cytometry showed that cultured cells were positive for CD90 and CD44, but negative for CD45.Results of immunohistochemistry demonstrated that vessel-derived BMSCs were negative for CD34, but positive for CD44. In cell disruption liquid of bFGF-transfected BMSCs, a significant positive zone of hybridization was visible at M, 23 000. However, no positive band was found in protein from pCDNA3.1(-) blank vector-transfected BMSCs.CONCLUSION: The bFGF gene was successfully transfected into BMSCs, and this target gene can stably express.
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This study sought to elucidate the effect of mechanical strain on the differentiation of mesenchymal stem cells into osteoblasts. Under the conditons of inducing osteoblasts, Immunohistochemical methods and RT-PCR technology were applied in osteogenic supplements medium to detect: (1) the expression of Alkaline phosphatase (ALP), Type I collagen (COL I ), Osterx (Osx) and Osteocalcin (OCN) mRNA, with cyclic strain (3%, 0.5 Hz) applied for 15 min, 30 min, 1 h, 2 h, 4 h, 3 d, 7 d, 14 d; (2) the expression of Osx mRNA and OCN mRNA with 3% strain for 1 h. The results showed: (1) ALP mRNA expression was higher at 7 days; COL I mRNA expression was greater obviously at 7 days and 14 days than that at 3 days and that of the unstrained cells; (2) the expression of Osx mRNA was up-regulated after 15min by strain stimulation,which was significantly increased at 30 min and 1 h in the unstrained cells. The expression of OCN mRNA was not affected in the unstrained cells at 15 min, whereas strain could promote the expression of OCN mRNA at this period. The expression of OCN mRNA was more obviously upregulated in the strained cells at 30 min and 1 h when compared with that in the unstrained cells; (3) the strain (1% and 3%) significantly promoted the expression of Osx mRNA; 10% strain had a little effect on Osx mRNA expression. The expression of OCN mRNA was up-regulated by 3% strain, whereas it had little effect at 1% and 10% strain. In summary, mechanical strain can promote the differentiation of mesenchymal stem cells into osteoblasts.