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1.
Chinese Journal of Primary Medicine and Pharmacy ; (12): 2353-2356, 2020.
Article Dans Chinois | WPRIM | ID: wpr-866609

Résumé

Objective:To explore the clinical effect of traditional Chinese medicine(TCM) fumigation-washing therapy combined with etofenamate cream wiping in the treatment of knee osteoarthritis.Methods:From September 2018 to April 2019, 176 cases of knee osteoarthritis were divided into two groups according to random number table method.The observation group (91 cases) was treated by etofenamate cream based on fumigation-washing therapy with TCM, while the control group (85 cases) was treated by etofenamate cream wiping only.Both two groups continued treatment for 2 weeks.The Lequesne score and effective rate of the two groups were achieved and analyzed.Results:At 1 d and 2 weeks after treatment, there were no statistically significant differences in the Lequesne score between the two groups(all P>0.05). After treatment for 1 month and 3 months, Lequesne scores of the observation group[(4.1±1.1)points, (4.6±1.0)points] were lower than those of the control group [(6.2±1.2)points, (7.5±1.4)points]( t=12.155, 15.598, all P<0.05). At 1 d and 2 weeks after treatment, there were no statistically significant difference in the effective rate between the two groups(all P>0.05). After treatment for 1 month and 3 months, the effective rates of the observation group were 63.7%(58/91) and 61.5%(56/91), respectively, which were higher than those of the control group [41.2%(35/85) and 18.8%(16/85)] (χ 2=8.98, 33.17, all P<0.05). Conclusion:Fumigation-washing therapy with TCM combined with etofenamate cream wiping has quick, lasting and safe effect in the treatment of knee osteoarthritis.

2.
Chinese Journal of Tissue Engineering Research ; (53): 3501-3507, 2016.
Article Dans Chinois | WPRIM | ID: wpr-492640

Résumé

BACKGROUND:Herba epimedi, a traditional Chinese medicine, has a long time in dealing with various orthopedic disorders. Icarinwithmany biological activites is one of the most important compositions of Herba epimedi. OBJECTIVE:Toinvestigate the effects of icarin on osteogenic differentiation of mesenchymal stem cels and the underlying mechanisms. METHODS:Bone marrow mesenchymal stem cels were treated using icarin with or without osteogenic mediumin vitro. Osteogenic differentiation markers, including runt-related transcription factor 2, osteocalcin and osterix, were detected by real time-qPCR. Alizarin red staining was used to measure calcium nodes generated by osteoblasts induced frombonemarrow mesenchymal stem cels. The proximal tibia bone structure of rats fed with icarin (2 mgperday) for 5 weeks was detected and analyzed by MicroCT. RESULTS AND CONCLUSION:Icarin was able to promote the expression of genes related to osteogenic differentiation in the absence or presence of osteogenic induction. Icarin could obviously increase the quantity of calcium nodes whenmesenchymal stem celswere cultured in the osteogenic medium. The animal experiment showed that icarin improved formation of trabecular bone.

3.
Chinese Journal of Hospital Administration ; (12): 123-125, 2016.
Article Dans Chinois | WPRIM | ID: wpr-487221

Résumé

Quality of care and safety are the lifeline of hospital performance and hospital management.With reference to the KTQ hospital quality certification system of Germany,Tongji Hospital built platforms to supervise outpatient,emergency,inpatient,surgical operation,nursing, hospital-acquired infection,and pharmacy management.By the connection and reaction of both online and offline systems,Tongji Hospital has built a systematic,informationized and precise medical quality and safety system for large public hospitals,safeguarding quality of care and safety of patients.

4.
Chinese Journal of Geriatrics ; (12): 665-667, 2014.
Article Dans Chinois | WPRIM | ID: wpr-450266

Résumé

Objective To observe the effects of different concentrations of glucose on the proliferation and differentiation of primary osteoblasts.Methods The identification of mouse primary osteoblasts was performed by alkaline phosphatase (ALP)staining and Von Kossa staining.Treating osteoblasts with different dose of glucose (5.5,15.5,25.5 mmol/L),the osteoblasts proliferation,ALP staining,and Runx2,OB markers ALP and OCN mRNA expression were observed.Real-time PCR was used for the determination of Runx2,OB markers ALP and OCN mRNA expression.Results With the increasing glucose concentrations,the osteoblasts cell proliferation was decreased.Compared with 5.5 mmol/L normal glucose,the ALP staining in 15.5 mmol/L group and 25.5 mmol/L group were decreased.The expressions were decreased by (36.7±6.2)% and (38.3±2.2)% in Runx2 mRNA,(26.7±7.2)% and (40.4±4.3)% in OCN mRNA respectively.ALP in 15.5 mmol/L group was reduced by (33.3±10.2)%,but increased by(50.8±10.4) % in 15.5 mmol/L group.Conclusions High glucose may decrease osteoblasts proliferation and activity,which may be one of the key pathogenesis factors of diabetic osteoporosis.

5.
The Journal of Practical Medicine ; (24): 548-551, 2014.
Article Dans Chinois | WPRIM | ID: wpr-446301

Résumé

Objective To compare the effects of sodium hyaluronate (SH) and celecoxib (CO) administration on the treatment of knee osteoarthritis (OA) and to investigate their influences on levels of uPA and MMP-3 in synovial fluid. Methods One hundred and thirty-six knee osteoarthritis (OA) patients from January 2010 to October 2011 were randomly enrolled into two groups: the SH group and the CO group. In the SH group, patients were injected with 2 mL sodium hyaluronate intra articulation once a week for 5 weeks. In the CO group , patients were given oral administration of celecoxib daily at a dosage of 200 mg for 5 weeks. Before and at 1 ,6 months after treatment, Lequesne′s index and VAS-pain were detected to assess the clinical results of these two drugs. The levels of uPA and MMP-3 in synovial fluid were measured by using ELISA assay. Results All patients were followed up for 6 months to 12 months. The Lequesne′s index and VAS-pain score were lowered at 1 and 6 moths after treatment in both the SH group and the CO group(P0.05). Dramatic reduction of the levels of uPA and MMP-3 in synovial fluid were observed in the SH group after treatment(P0.05). Conclusion The redueced levels of uPA and MMP-3 in synovial fluid after treatment of sodium hyaluronate may contribute to its longer-lasting effect than that of celecoxib. Therefore , the combination of sodium hyaluronate with celecoxib may lead to better therapeutic effect on OA patients.

6.
Journal of Jilin University(Medicine Edition) ; (6): 650-654, 2014.
Article Dans Chinois | WPRIM | ID: wpr-491221

Résumé

Objective To detect the expression levels of urokinase-type plasminogen activator (uPA), matrix metalloproteinase-3 (MMP-3),MMP-9,MMP-13 and MMP-14 in the patients with osteoarthritis(OA)before and after arthroscopic debridement,and to explore the influence of arthroscopic debridement in the expressions of uPA, MMP-3,MMP-9,MMP-13, and MMP-14.Methods 420 cases of synovial fluid from knee OA patients undergoing arthroscopic debridement were obtained before operation. After six months follow-up, 350 cases of synovial fluid samples were obtained and according to inclusion and exclusion criteria, 228 synovial fluid were selected to analyze.The expression levels of uPA,MMP-3,MMP-9,MMP-13,and MMP-14 were measured by ELISA assay.Pain intensity of these patients before operation and six months after operation were recorded using the Visual Analogue Scale/Score(VAS).The differences of the expression levels of uPA,MMP-3,MMP-9, MMP-13,and MMP-14 between before operation and after operation were compared.The relationship between the expression levels of uPA, and MMP-3, MMP-9, MMP-13, MMP-14 and VAS was analyzed with Spearman analysis.Results All the patients were followed up for 36.5 months. Compared with before operation, the expression levels of uPA and MMP-3 in the synovial fluid of the patients after arthroscopic debridement were significantly decreased(P<0.01),the expression levels of MMP-9 and MMP-13 were also decreased (P<0.05), but the MMP-14 expression level showed no significant change.The expression levels of uPA,MMP-3,MMP-9, MMP-13,MMP-14 were positively associated with VAS before arthroscopic debridement (r=0.361,r=0.417, r=0.136,r=0.514,r=0.156,P<0.05 );uPA and MMP-3 were positively correlated with VAS after arthroscopic debridement(r=0.981,r=0.831,P<0.01),as well as the expression level of MMP-13 and VAS, but there were no significant differences between the expression levels of MMP-9, MMP-14 and VAS. Conclusion The decreased levels of uPA,MMP-3 and MMP-13 in synovial fluid may contribute to the pain-relief effects of arthroscopic debridement.

7.
Chinese Journal of Physical Medicine and Rehabilitation ; (12): 33-36, 2013.
Article Dans Chinois | WPRIM | ID: wpr-432342

Résumé

Objective To observe the effect of early rehabilitation therapy on recovery from reoperation for recurrent lumbar disc herniation (RLDH).Methods Sixty-five cases who received surgery for RLDH between 2007 and 2009 were randomly divided into a rehabilitation group and control group.Both groups were treated with the same surgical approach and routine treatment.Early and comprehensive rehabilitation therapy was provided in the rehabilitation group during the perioperative period,including preoperative and postoperative muscle strength training,postoperative sitting and standing balance training,and acupuncture.The control group was instructed only in general exercise.Before the operation and 2 weeks and 3,6,12 and 24 months afterward,the surgical outcomes of all cases were assessed using the JOA score and the improvement rate in the JOA score.Any postoperative complications and intervertebral fusion were also observed.Results The average postoperative JOA scores of both groups were significantly higher than their preoperative scores.At all of the time points after the operation,the average JOA scores and all improvement rates in the rehabilitation group were significantly higher than those in the control group.Postoperative complications such as deep venous thrombosis,urinary retention and constipation were significantly less among the rehabilitation group than among the controls.All the intervertebral bone implants were well fused on time.Conclusion Early rehabilitation can significantly improve the effectiveness of RLDH reoperation and reduce the incidence of postoperative complications.It is recommended for clinical application.

8.
Chinese Journal of Physical Medicine and Rehabilitation ; (12): 172-176, 2012.
Article Dans Chinois | WPRIM | ID: wpr-419730

Résumé

Objective To investigate the effect of 620 nm red light on chondrogenic differentiation in rat precartilaginous stem cells (PSCs). Methods Rats' PSCs were isolated and purified using magnetically activated cell sorting and cultured in vitro.The PSCs were exposed once to 620 nm wavelength red light from a light-emitting diode (LED) with an irradiation energy of 0.5 J/cm2,1 J/cm2,2 J/cm2 or 4 J/cm2.Any effect was confirmed by Alcian blue staining,immunohistochemistry and observing histomorphological changes under a light microscope,as well as detection using a reverse transcription polymerase chain reaction (RT-PCR). Results After being induced for 14 d,the PSCs exhibited polygonal and round shapes. Alcian blue and type Ⅱ collagen immunohistoehemistry staining showed positive results,but the control group had no significant change.RT-PCR showed that the mRNA expression of Sox9 and type Ⅱ collagen increased significantly compared with the control group. Conclusion Low energy 620 nm red light can enhance chondrogenic differentiation in PSCs significantly.

9.
Chinese Journal of Physical Medicine and Rehabilitation ; (12): 178-181, 2012.
Article Dans Chinois | WPRIM | ID: wpr-428496

Résumé

Objective To study the impacts of dynamic compressive stress on the mRNA expression of osteopontin ( OPN ),runt related gene 2 ( Runx2 ),osteocalcin ( OC ),osterix,alkaline phosphatase (ALP) and bone morphogenetic protein 2 (BMP-2) in the osteoblasts of Sprague-Dawley (SD) rats. Methods Osteoblasts extracted from skull periosteum tissue of neonatal SD rats were digested using trypsin and collagenase (Ⅰ),then were subcultured and amplified in vitro.ALP staining and alizarin red staining were performed to identify the purified cells.The cells were treated with compressive stress at 20,50 or 100 mmHg for 24 h.The expression levels of OPN,Runx-2,OC,osterix,ALP and BMP-2 were measured and quantitatively analysed using a real-time quantitative polymerase chain reaction. Results Under 20 mmHg of dynamic compressive stress the expression levels of OPN,Runx2,OC,osterix,ALP and BMP-2 all were elevated compared with the control group.The peak expression oecured under 50 mmHg pressure. The expression levels did not change significantly compared with the control group under 100 mmHg pressure. Conclusions Moderate dynamic compressive stress can promote the expression of OPN,Runx-2,OC,osterix,ALP and BMP-2 mRNA in osteoblasts,which might be an important mechanism for promoting the union of fractures.

10.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 82-6, 2012.
Article Dans Anglais | WPRIM | ID: wpr-636045

Résumé

Matrix metalloproteinase-2 (MMP-2) level and the ERK1/2 signal pathway are dependent factors for the growth and metastasis of cancer. However, the impact of MMP-2 in combination with ERK1/2 in tumor patients with drug resistance is unknown. To determine the relationship between MMP-2 and the ERK1/2 signal pathway, we established an adriamycin (ADM)-induced MG-63 (ADM-MG-63) cell line. With the increase of the ERK1/2 pathway blocker PD98059, we detected the expression levels of MMP-2 and p-ERK1/2 by Western blot in ADM-MG-63 cells. In ADM-MG-63 cells transfected with MMP-2-siRNA, the expression of ERK1/2 was detected for understanding the function of the ERK1/2 signal pathway. Three siRNAs for MMP-2 (MMP-2-siRNA) were designed, and the optimal one was selected and tested at different time points of 24, 48 and 72 h. Under an ADM-induced condition, ADM-MG-63 cells were finally stable living in the medium of ADM (200 ng/mL). PD98059 could effectively suppress the expression levels of p-ERK1/2 and MMP-2. When the MMP-2 was silenced by using MMP-2-siRNA, the expression of p-ERK1/2 was enhanced. It is concluded that MMP-2 may be involved in ADM resistance dependent on ERK1/2 signal pathway, suggesting interference in ERK1/2 may be a new method of targeted therapy for tumor resistance.

11.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 75-81, 2012.
Article Dans Anglais | WPRIM | ID: wpr-636044

Résumé

The osteogenic in vitro effect of low intensity pulsed ultrasound (LIPUS) on SD rat adipose-derived stem cells (ADSCs) was investigated. Rat ADSCs underwent LIPUS (intensity=100 mW/cm(2)) or sham exposure for 8 min per treatment once everyday in vitro, and then the alkaline phosphatase (ALP) activity and mineralized nodule formation were assessed to evaluate the osteogenic effect of LIPUS on ADSCs. To further explore the underlying mechanism, the osteogenic-related gene mRNA expression was determined by using reverse transcriptase-polymerase chain reaction (RT-PCR) at 1st, 3rd, 5th, 7th day after exposure repectively. Westen blot was used to evaluate the protein expression levels of two osteogenic differentiation associated genes at 7th and 14th day repectively. It was found that ALP activity was increased after LIPUS exposure and LIPUS resulted in mineralized nodule formation of ADSCs in vitro. LIPUS-treated ADSCs displayed higher mRNA expression levels of runt-related transcription factor 2 (Runx2), osteocalcin (OCN), ALP and bone sialoprotein (BSP) genes than controls, and the protein levels of Runx2 and BSP were also increased. The results suggested that LIPUS may induce the osteogenic differentiation of ADSCs in vitro.

12.
Chinese Journal of Trauma ; (12): 659-662, 2011.
Article Dans Chinois | WPRIM | ID: wpr-416460

Résumé

Objective To study the causes and theoretical basis for good bone healing ability of magnetic Porous Ca3 (PO4) 2 ( MPTCP). Methods Seven MPTCP specimens with size of 2 cm × 1 cm × 0.5 cm were placed in the material physical system for detecting 42 times and the mean detection value was used to measure the MPTCP curve. The attachment 16451B of impedance spectrometer HP RLC was employed to measure dielectric spectroscopy and dielectric spectroscopy of MPTCP. Four-wire method was used to measure the impedance of MPTCP. Results The magnetic intensity changed rapidly when magnetic field was in a range of-10,000-10,000 Oe. The peak of dielectric spectroscopy and impedance of magnetic bioceramics was in the range of 103-104 Hz. When the external electromagnetic wave of frequency was ≤ 1 000 Hz, electrical impedance of MPTCP was large;while when the electromagnetic wave frequency was≥1 000 Hz, the impedance was relatively small and stable. Conclusion The environmental magnetic fields may change the magnetic and electric behavior of MPTCP and promote the biological healing, which may be the cause for the good bone healing ability of MPTCP.

13.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 524-9, 2011.
Article Dans Anglais | WPRIM | ID: wpr-635424

Résumé

This study examined the construction of eukaryotic expression plasmid of human transforming growth factor-β3 (hTGF-β3) and its inducing effect on the differentiation of precartilaginous stem cells (PSCs) into chondroblasts. hTGF-β3 gene was amplified by using polymerase chain reaction (PCR) and then inserted into the eukaryotic expression plasmid pcDNA3.1 to construct the eukaryotic expression plasmid pcDNA3.1(+)-hTGF-β3. Rat PSCs were isolated and purified by employing an immunomagnetic cell sorting system. pcDNA3.1(+)-hTGF-β3 was transfected into purified PSCs with the use of linear polyamines. The expression of TGF-β3 and cartilage-specific extracellular matrix (ECM) components was detected after transfection by real-time quantitative PCR, ELISA, immunochemistry and Western blotting, respectively. The results showed that the eukaryotic expression plasmid pcDNA3.1(+)-hTGF-β3 was successfully established as identified by enzyme digestion and DNA sequencing. Real-time quantitative PCR and ELISA revealed that hTGF-β3 was strongly expressed in pcDNA3.1(+)-hTGF-β3-transfected PSCs. Real-time quantitative PCR, immunochemistry and Western blotting showed that the cartilage-specific ECM markers, i.e., cartilage oligomeric matrix protein (COMP), Aggrecan, collagen type X and II were intensely expressed in the pcDNA3.1(+)-hTGF-β3-transfected cells. It was concluded that hTGF-β3 could be stably expressed in pcDNA3.1(+)-hTGF-β3-transfected PSCs and induce the differentiation of PSCs into chondroblasts.

14.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 353-8, 2011.
Article Dans Anglais | WPRIM | ID: wpr-635114

Résumé

This study examined the effect of small interfering RNA-mediated β-catenin knockdown on the survival, invasion and chemosensitivity of human osteosarcoma cells (U2-OS cells). The siRNA against β-catenin was constructed and transfected into U2-OS cells. The expression of β-catenin was detected by qRT-PCR and Western blotting. Cell growth and apoptosis was detected in the presence or absence of doxorubicin by MTT and flow cytometry, respectively. Cell invasion ability was measured by transwell assay. The results showed that the transfection of β-catenin siRNA resulted in decreased expression of β-catenin, suppression of invasion and motility of U2-OS cells, reduced chemosensitivity to doxorubicin in vitro, and little change in cell growth and apoptosis. Additionally, down-regulated MT1-MMP expression was found after transfection. It was concluded that knockdown of β-catenin gene may decrease the invasive ability of human osteosarcoma cells through down-regulated MT1-MMP expression, and the chemosensitivity of osteosarcoma cells against doxorubicin.

15.
Chinese Journal of Orthopaedics ; (12): 213-218, 2011.
Article Dans Chinois | WPRIM | ID: wpr-384272

Résumé

Objective To assess the feasibility and clinical results of video-assisted high anterior transcervical approach (Smith-Robinson) in treatment of spinal lesions of the craniovertebral junction. Methods Between April 2007 to October 2009, nineteen consecutive patients with spinal lesions of the craniovertebral junction were included in the study. There were 9 males and 10 females aged from 16 to 62 years old with a mean of 32 years. The primary pathologies included 4 cases with chronic odontiod fracture, 2 cases with purely irreducible atlantoaxial dislocation, 6 cases with os odonteideum, 1 case with Marfan synd rome, 1 case with primary basilar invagination from Kippel-Feil syndrome, 3 case with axis tumor and 1 case with irreducible rheumatoid atlantoaxial dislocation. All of the patients underwent combined video-assisted high anterior transcervical procedure and posterior fixation at one-stage. The anterior procedure included atlantoaxil release and reduction (8 cases), odontoidectomy (8 cases), and intralesional extracapsular excision and reconstruction (3 tumor cases). The posterior technique were C1-C2 pedicle screw fixation (13 cases), C1-C3 pedicle screw fixation (2 cases), and occipitalcervical fusion (4 cases). Results Anatomical reduction was achieved in eight cases with anterior release and reduction. Tumors were completely removed in three cases with axial tumor. The mean follow-up was 14 months (6-36 months). All of them achieved solid bone fusion. In the 14 patients with symptoms of spinal cord dysfunction, the average Japanese Orthopaedic Association (JOA)score had improved from 9.1±3.3 preoperatively to 14.1±2.9 postperatively. The improvement rate was excellent for 7 cases, good for 5 cases, fair for lcase and poor for 1 case. One patient experienced leakage of cerebrospinal fluid which was resolved by bioprotein gelatin blocking and lumbar subarachnoid continuous drainage within 1 week. Dysphagia which occurred in 3 cases responded well to dexamethason and mannitol.No infection and hardware failure were observed. Conclusion Video-assisted high anterior transcervical procedure is a safe and effective alternative for treating spinal lesions in the craniovertebral junction.

16.
Chinese Journal of Cancer Biotherapy ; (6): 71-76, 2010.
Article Dans Chinois | WPRIM | ID: wpr-404246

Résumé

Objective: To investigate the differentially expressed genes in osteosarcoma cell lines with various metastatic potentialities, and to screen for new candidate genes related to metastasis of osteosarcomas. Methods: The total RNAs of a lowly metastatic and a highly metastatic osteosarcoma cell lines (M6 and M8) were extracted. Differentially expressed genes in the two osteosarcoma cell lines were studied by cDNA microarray. The hybridization signals were scanned with a Generation Ⅲ array scanner and analyzed by Imagequant 5.0 software. Typical differentially expressed genes were further verified by real-time quantitative PCR. Results: There were 330 differentially expressed genes between M6 and M8 cells. In the high-metastasis M8 cells, 178 genes were up-regulated and 152 genes were down-regulated compared to the low-metastasis M6 cells, with 43 extremely up-regulated and 49 extremely down-regulated. The differentially expressed genes were mainly associated with cell proliferation, indicating these genes might be related to the inhibition of M6 cells. Other differentially expressed genes included those associated with the regulation of gene expression and signal transduction, indicating these genes might be correlated with tumor metastasis. Conclusion: cDNA microarray shows an advantage in identifying genes associated with metastasis of osteosarcoma. In M8 subset of MG63 osteosarcoma cells,43 genes are up-regulated and 49 genes are down-regulated, which may be related with metastasis of osteosarcoma.

17.
Chinese Journal of Tissue Engineering Research ; (53): 1009-1014, 2010.
Article Dans Chinois | WPRIM | ID: wpr-403562

Résumé

BACKGROUND: Models concerning tumor external environment mainly concentrated on laboratory two-dimensional culture and in vitro animal experiment, which lack of three-dimensional stereo.OBJECTIVE: To establish in vitro bone metastasis stereo models of human prostate carcinoma, and to investigate the effect of stem cells on proliferation rate and clustering size of prostate carcinoma cells. METHODS: Bone marrow mesenchymal stem cells (BMMSCs) were extracted from 2 clean grade SD rats. Alginate was used to simulate medullary microenvironment, where prostate carcinoma cells and BMMSCs were co-culturedd. Growth of the cells in the three-dimensional model was observed through microscope and histological sections. The carcinoma cells were transfected with green fluorescent protein. The proliferation of monoclonal cells clustering was observed under light microscope and fluorescence microscope. RESULTS AND CONCLUSION: In the co-culture group, the clustering speed, clustering amount and tumor formation rate were greater that those of the control group. The monoclonal cells clustering was formed at 7.75 days and 6.00 days in the control and co-culture groups, respectively, with cell counts of (95.13±11.63) and (112.53±14.67) after 10 days. The formation rate of fluorescent cell clones was (77.10±6.85)% in the control group and (64.55±6.21)% in the co-culture group, the difference had significance. The results suggested that: the alginate microenvironment is conductive to proliferation and clustering of prostate carcinoma cells and BMMSCs.

18.
Chinese Journal of Tissue Engineering Research ; (53): 223-226, 2010.
Article Dans Chinois | WPRIM | ID: wpr-403410

Résumé

BACKGROUND: The precartilaginous stern cells are limited regarding in vitro proliferative capacity, but the immortalized cell lines can provide a large number of stable immortalized cells, and simian virus 40 large T antigen gene (SV40Tag) is one of gene fragments which are commonly used and effective in vitro immortalized ceils. OBJECTIVE: To construct human immortalized precartilaginous stem cells (IPSCs) using human precartilaginous stem calls induced by SV40LTAg gane. METHODS: The human immortalized precartilaginous stem calls were isolated from aborted fetus and purified with enzyme digestion and immunomagnetic beads screening method. By using liposome-mediated gene transfection technology, plasmid pCMVSV40T/PUR containing SV40Tag was transfected in primary embryonic precartilaginous stem cells, while non-transfected cells sewed as negative controls. Positive clones were cultured to observe the cell morphology and the passage recovery, to calculate cell survival rata and population doubling time, to drew call growth curve. Immunofluorescence cytochemistry was used to detect the expression of IPSCs fibroblast growth factor receptor 3, the expressions of SV40Tag and fibroblast growth factor receptor 3 in the human precartilaginous stem cells were determined by RT-PCR. RESULTS AND CONCLUSION: Morphology of human IPSCs seemed coincidence with primary human precartilaginous stem cells. The survival rate of human IPSCs was not influenced by subculture, freezing and recovery, but the survival rate was descended in the human precartilaginous stem cells at the 6~(th) and 10~(th) passages (P < 0.01). Compared with cells at the 6~(th) and 10~(th) passages, the proliferation of human IPSCs was greater, with short population doubling time and high growth rate (P < 0.01). The immunofluorescence showed that fibroblast growth factor receptor 3 was positive in human IPSCs at the second passage, and the RT-PCR results of fibroblast growth factor receptor 3 revealed a specific amplification band at 400 bp,.while that of SV40Tag revealed at 560 bp. No band was seen in the primary cells. It is indicated that SV40Tag human IPSCs can be constructed successfully using immunomagnatic bead screening technology and liposome transfection technique.

19.
Chinese Journal of Tissue Engineering Research ; (53): 726-731, 2010.
Article Dans Chinois | WPRIM | ID: wpr-402902

Résumé

BACKGROUND: Multilevel cervical disc herniation accompanying ossification of posterior longitudinal ligament (OPLL) can seriously hurt the spinal cord. Which way of operative approach is more preferable is still uncertain. Whether one-stage combined anterior and posterior operation can achieve better effects remains unclear.OBJECTIVE: To explore the therapeutic effect of one-stage anterior and posterior operation for the treatment of multilevel cervical disc herniation accompanying OPLLMETHODS: Seventeen patients with multilevel cervical disc herniation and OPLL were treated by one-stage anterior and posterior operation at the Department of Orthopaedics, Tongji Hospital, were selected, including 11 males and 6 females, aged 42-74 years,mean aged 51.5 years. X-ray film, CT or MRI before operation showed the cervical cord was compressed by the multilevel herniated cervical discs or the ossified posterior longitudinal ligament. The stability and fusion of the injured segments were observed by regular postoperative X-ray film.RESULTS AND CONCLUSION: All patients were received a 6-36 months follow-up (24.5 months on average). The postoperative JOA scores were (12.88±2.47) points, which was greater than that of preoperatively [(6.41±1.28) points, P<0.05]. The improvement-rate of the spinal function after 6 months included 5 excellent cases, 7 good cases, and 4 fair cases. The excellent-good rate was 71%. All the patients got completely reduction, and all grafts got fused at 3-4 months after operation. The cervical intervertebral height and lordosis were satisfied maintained and there was no complication related to internal fixation breakage, loosening or displacement. It suggested that one-stage anterior and posterior operation can provide satisfied decompression earlier and rebuild the spinal stability in time, which is a safe and effective surgical intervention for multilevel cervical spondylotic myelopathy accompanying OPLL.

20.
Chinese Journal of Tissue Engineering Research ; (53): 1892-1896, 2010.
Article Dans Chinois | WPRIM | ID: wpr-402538

Résumé

BACKGROUND:Schwann cell is one of the major seed cells In peripheral nervous system and plays an important role in neural injury and neural disease.However,the source of Schwann cells is limited.And the purity of Schwann cells is affected due to the pollution of fibroblasts.Many purified methods have been proposed,but every one has its defect to satisfy the clinical demand.OBJECTIVE:To compare the differences among differential adhesion purified method,cold jet purified method,immunomagnetic beads selection purified method and G418 selection purified method to purify Schwann cells of neonatal rat in vitro.METHODS:Bilateral sciatic nerves of SD rats were harvested under sterile condition.Schwann cells were purified respectively using differential adhesion purified method,cold jet purified method,immunomagnetic beads selection purified method and G418 selection purified method.Cell viability was compared,and cell purity was determined by immunohistochemistry.RESULTS AND CONCLUSION:The purity of Schwann cells separated by differential adhesion method was low,but the viability was fair.The purity and viability of cells following cold jet method immunomagnetic beads selection method was high.The purity of cells separated by immunomagnetic beads selection methods was similar to that of cold jet method immunomagnetic beads selection method,but the cell viability was worse.The cell viability following G418 selection method was bad,but the purity was high.

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