Targeted binding of estradiol with ESR1 promotes proliferation of human chondrocytes by inhibiting activation of ERK signaling pathway / 南方医科大学学报

OBJECTIVE@#To investigate the effect of estradiol (E2)/estrogen receptor 1 (ESR1) on the proliferation of human chondrocytes and explore the molecular mechanism.@*METHODS@#The Ad-Easy adenovirus packaging system was used to construct and package the ESR1-overexpressing adenovirus Ad-ESR1. Western blotting and qPCR were used to detect the expression of ESR1 protein and mRNA in human chondrocyte C28I2 cells. In the cells treated with different adenoviruses, the effects of E2 were tested on the expressions of proteins related with cell autophagy and apoptosis and the phosphorylation of ERK signaling pathway using Western blotting. Immunofluorescence assay was used to observe the intracellular autophagic flow, flow cytometry was performed to analyze the cell apoptosis rate and the cell cycle changes, and qPCR was used to detect the expressions of PCNA, cyclin B1 and cyclin D1 mRNAs. The inhibitory effect of the specific inhibitor of ERK on the expressions of autophagy- and apoptosis-related genes at both the protein and mRNA levels were detected using Western blotting and qPCR.@*RESULTS@#Transfection with the recombinant adenovirus overexpressing ESR1 and E2 treatment of C28I2 cells significantly enhanced the expressions of autophagy-related proteins LC3, ATG7, promoted the colocalization of LC3 and LAMP1 in the cytoplasm, increased the expressions of the proliferation-related marker genes PCNA, cyclin B1 and cyclin D1, and supressed the expressions of cleaved caspase-3, caspase-12 and pERK. RNA interference of ESR1 obviously lowered the expression levels of autophagy-related proteins in C28I2 cells, causing also suppression of the autophagic flow, increments of the expressions of apoptosis-related proteins and pERK, and down-regulated the expressions of the proliferation marker genes. Blocking ERK activation with the ERK inhibitor obviously inhibited the effects of E2/ESR1 on autophagy, proliferationrelated gene expressions and cell apoptosis.@*CONCLUSIONS@#The targeted binding of E2 with ESR1 promotes the proliferation of human chondrocytes possibly by inhibiting the activation of ERK signaling pathway to promote cell autophagy and induce cell apoptosis.

Comparison of different anesthesia techniques on early prognosis in patients undergoing hip joint re-placemen / 中华麻醉学杂志

Objective To compare the effects of different anesthesia techniques on early prognosis in patients undergoing hip joint replacement. Methods The demographic, preoperative and postoperative data of 478 patients, aged 18-95 yr, of American Society of Anesthesiologists physical statusⅠ-Ⅳ, who underwent elective unilateral hip joint replacement in Tongji Hospital from May 2014 to December 2016, were retrospectively analyzed. Patients were divided into general anesthesia group (group GA, n=197), peripheral nerve block group ( group PNB, n=147) and peripheral nerve block combined with general an-esthesia group ( group PNB+GA, n=134) . The amount of crystalloid solution and colloid solution infused, consumption of sufentanil and requirement for vasoactive agents were recorded during operation. The dura-tion of anesthetic recovery room stay, length of hospital stay before and after operation and total length of hospital stay were recorded. The development of complications within 48 h after operation, therapy after ad-mission to intensive care unit and in-hospital fatality were also recorded. Results Compared with group GA, the intraoperative consumption of sufentanil was significantly decreased in group PNB+GA, and the a-mount of crystalloid solution infused, urine output, consumption of sufentanil, requirement for vasoactive agents and incidence of postoperative hypoxemia, pulmonary infection and acute cerebral infarction were significantly decreased in group PNB+GA ( P<0. 05) . Compared with group PNB+GA, the consumption of sufentanil, requirement for vasoactive agents and incidence of postoperative hypoxemia, pulmonary infec-tion and acute cerebral infarction were significantly decreased in group PNB (P<0. 05). Conclusion Compared with general anesthesia or with peripheral nerve block-general anesthesia, peripheral nerve block is more helpful in improving early prognosis in patients undergoing hip joint replacement.

Differential regulation of SREBP 1 c/cm on transcriptional activity and expression of PERK promoter / 基础医学与临床

Objective To investigate the effect of sterol regulating element binding protein (SREBP1c) and its ac-tive form (SREBP1cm) on human protein kinase R-like endoplasmic reticulum kinase (PERK).Methods Re-porter victors of PERK promoter and its truncations were constructed with pGL 3-basic and co-transfected with internal reference pRL-SV40 into cell and luciferase activity was detected .pcDNA3.1 ( -)-SREBP1c or pcDNA3.1 ( -)-SREBP1cm was co-transfected with PERK promoter transcriptional activity core regions and the detection of dual -lu-ciferase reporter gene was used to analyze the regulation of SREBP 1c/1cm on PERK promoter transcriptional activity . The expression level of PERK mRNA and protein were detected by RT-PCR and Western blot .Results PERK pro-moter and truncations were successfully constructed into pGL 3-basic, and PERK promoter core area of transcription-al activity had determined;Dual-luciferase report gene showed that SREBP 1c inhibited PERK promoter transcrip-tional activity and SREBP1cm promoted PERK promoter transcriptional activity .RT-PCR and Western blot showed that SREBP1c decreased PERK mRNA and protein expression , but SREBP1cm increased PERK mRNA and protein expression, which was consistent with the detection of dual-luciferase report gene .Conclusions SREBP1c and SREBP1cm have a opposite regulation effect on PERK promoter transcriptional activity and its expression .

Experimental study of osteogenic differentiation of precartilaginous stem cells induced by calcitonin gene-re-lated peptide / 中华物理医学与康复杂志

Objective To investigate the effect of calcitonin gene-related peptide ( CGRP ) in inducing os-teogenic differentiation of rat precartilaginous stem cells in vitro and the underlying mechanisms. Methods Rat pre-cartilaginous stem cells ( PSCs) were cultured in complete osteogenesis medium containing DMEM/F-12 medium and different concentrations (0, 10-8,10-9,10-10mol/L) of CGRP, the morphology changes of PSCs were observed. The proliferation of PSCs was examined at different time points by CCK-8. All the PSCs were then randomly assigned to an experimental group and a control group. The PSCs in the experimental group were cultured in complete osteogenesis medium with 10-10 mol/L CGRP , while the control group cultured merely in complete osteogenesis medium was re-ceived no special intervention. Both groups were stained by Alizarin Red and the expression of alkaline phosphatase (ALP) was detected. The osteogenic genes (RUNX2,OPN and BGP) were measured by use of RT-PCR. The activa-tion of Wnt/β-catenin signaling pathway was tested by using Western blotting to evaluate the effect of CGRP . Results Compared to the control group ( the concentration of CGRP was 0 mol/L) , the concentration of ALP was significantly higher in the experimental group, calcium deposition was significantly more obvious, and the expression of the osteogenic genes such as RUNX2,OPN and BGP as well as theβ-catenin protein expression were up-regulated significantly. However, CGRP had no effect on cell proliferation. Conclusion CGRP activated Wnt/β-catenin sig-nal pathway and induced osteogenic differentiation of precartilaginous stem cells.

Icariin promotes osteogenic differentiation of bone marrow mesenchymal stem cells / 中国组织工程研究

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.

Effects of high glucose on the proliferation and differentiation of primary osteoblasts / 中华老年医学杂志

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.

Effects of Sox-9 gene silenced by small interfering RNA on proliferation and apoptosis of epiphysis stem cells / 中国组织工程研究

BACKGROUND:Current research has shown that many cellgrowth factors can promote the proliferation and differentiation of epiphysis stem cells, and this regulation is closely related to Sox-9. OBJECTIVE:To investigate the effects of smal interfering RNA-induced specific silence of Sox-9 gene on the proliferation and apoptosis of epiphysis stem cells. METHODS:Epiphysis stem cells were isolated from the ring of La Croix with enzyme digestion and purified with magnetic activated cellsorting, identified by immunocytochemistry assay. The cells in good conditions were selected to be transfected by Sox-9 silenced by smal interfering RNA with fluorescent marker, and then were observed under the fluorescent microscope to check the transfection efficiency. Next step, epiphysis stem cells were divided into 2 groups:group one was cultured normal y as control group;group two was transfected by Sox-9 silenced by smal interfering RNA through LipofectamineTM 2000 as experimental group. RESULTS AND CONCLUSION:The primary epiphysis stem cells were separated and purified, which were of stable growth and tightly attachment. The results of immunohistochemistry and immunofluorescence showed the epiphysis stem cells expressed cel-specific markers, fibroblast growth factor receptor 3. After transfection, reverse transcription-PCR results showed that Sox-9 gene expression in epiphysis stem cellwas inhibited specifical y;3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide results showed that cellproliferation in experimental group decreased significantly compared with the control group (P<0.05), and the cellapoptosis detected by flow cytometry showed that the experimental group increased as compared with the control group (P<0.05). Sox-9 gene plays an important role in regulating the proliferation and apoptosis of rat epiphysis stem cells.

A genetic study of dynamic compressive stress promoting proliferation and differentiation of rat osteoblasts / 中华物理医学与康复杂志

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.

Enhancing chondrogenic differentiation in precartilaginous stem cells with 620 nm red light / 中华物理医学与康复杂志

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.

Implication of expression of Nanog in prostate cancer cells and their stem cells / 华中科技大学学报（医学）（英德文版）

Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells. The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells, and the relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells. By using magnetic active cell sorting (MACS), we isolated a population of CD44(+)/CD133(+) prostate cancer cells that display stem cell characteristics from PC3 cell line. Immunohistochemistry revealed positive expressions of CD44, CD133 and α(2)β(1)-integin in the isolated cells. CCK-8 analysis showed that isolated cells had a strong proliferative ability. The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing, indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line. Western blotting exhibited that the isolated cells had higher experession of Nanog, an embryonic stem marker, as compared with PC3 cells. Our study showed that Nanog might be helpful in sustaining the self-renewal and the undifferentiation of prostate cancer stem cells, and may serve as a marker for prostate cancer stem cells for isolation and identification.

The effects of rational emotive behavior therapy in the rehabilitation ot patients with spinal cord injury / 中华物理医学与康复杂志

Objective To study the effects of rational emotive behavior therapy (REBT) in the rehabilitation of patients with spinal cord injury (SCI).MethodsA total of 300 SCI patients from six institutions were divided into a research group and a control group. Systematic rehabilitation was given to the patients of the control group, while systematic rehabilitation and REBT were given to the patients of the research group. All patients were followed up for 1 year. Psychological state, activities of daily living (ADL), and quality of life (QOL) were evaluated with a symptom check list (SCL-90), a modified version of the Barthel index (MBI) and the World Health Organization's quality of life assessment (WHOQOL-100).ResultsOne year after treatment, the improvement in QOL of patients in the research group was better, on average, than that in the control group. The differences were primarily in mental items. The ADL ability of patients in both groups improved, and any differences were not significant. The psychological state of patients in the research group had improved significantly 1 year after treatment, while the improvements in the control group were not significant on average, except in terms of interpersonal sensitivity.ConclusionREBT had little effect on the ADL ability of SCI patients, however, it improved their psychological state significantly, and thus improved their QOL.

Screening of differentially expressed genes in osteosarcoma cell lines with various metastatic potentialities / 中国肿瘤生物治疗杂志

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.

Co-culture of prostate carcinoma cells with alginate and bone marrow mesenchymal stem cells: To observe the effect of stem cells on proliferation speed and clustering size of prostate carcinoma cells / 中国组织工程研究

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.

Construction of immortalized human precartilaginous stem cell lines / 中国组织工程研究

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.

One-stage anterior and posterior operation for multilevel cervical disc herniation accompanying ossification of posterior longitudinal ligament / 中国组织工程研究

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.

Schwann cells purification by four different methods in vitro / 中国组织工程研究

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.

The effects of high-intensity pulsed electromagnetic field on proliferation and differentiation of neural stem cells of neonatal rats in vitro / 华中科技大学学报（医学）（英德文版）

The effects of high-intensity pulsed electromagnetic stimulation (HIPEMS) on proliferation and differentiation of neonatal rat neural stem cells in vitro were investigated. Neural stem cells derived from neonatal rats were exposed to 0.1 Hz, 0.5-10 Tesla (T) [8 groups of B-I, respectively], 5 stimuli of HIPEMF. The sham exposure controls were correspondingly established. Inverted phase contrast microscope was used to observe the cultured cells, MTT assay to detect the viability of the cells as expressed by absorbance (A) value, and flow cytometry to measure differentiation of neural stem cells. The results showed that A values of neural stem cells in both 3.0 T and 4.0 T groups were significantly higher than the other groups 24 to 168 h post HPEMS, indicating a strong promotion of the growth of neural stem cells (P0.05). It was suggested that 0.1 Hz, 5 pulses stimulation of HPEMS within certain scale of intensity (0.5-10.0 T), significantly promoted the growth of neural stem cells with the rational intensity being 4.0 T.

Combining physiotherapy with ballon kyphoplasty to treat osteoporotic vertebral compression fracture / 中华物理医学与康复杂志

Objective To evaluate the safety and effectiveness of balloon kyphoplasty combined with physiotherapy in the treatment of osteoporotic vetcrbral compression fractures (OVCFs). Methods A retrospective review was of 12 OVCF cases ( including 16 fracture vertebrae) treated with balloon kyphoplasty was performed. Each patient had also been treated with anti-osteoporotic medication and the Rehabilitation of Osteoporosis Program-Exercise (ROPE) protocol, and each had received a six-month follow-up visit. The following parameters were recorded before the operation and 1 day and 6 months afterward : fracture recurrence, the severity of pain before and after treatment, and the change of vertebral height. The severity of pain was evaluated by using a visual analogue scale (VAS). Results During the treatment, no complication or adverse event was found. The average VAS values preoperation, postoperation and at the 6-month follow-up were 7.6±1.3, 2.1±1.2 and 2.6±1.4, respectively. The average changes in vertebral height preoperation, postoperation and at the 6-month follow-up were 49.2±18% , 74.3 ±14% and 69.8±16% respectively. All of the measures evaluated improved noticeably after the combined treatment. Conclusion Balloon kyphoplasty combined with anti-osteoporotie medication and physiotherapy showed good effects in the treatment of osteoporotic veterbral compression fractures.

Separation,identification and immortalization of precartilaginous stem cells from neonatal rats / 中国组织工程研究

BACKGROUND: Precartilaginous stem cells (PSCs) have strong proliferation ability and differentiation potential,but they are instable and prone to differentiate.Importing exogenous gene could immortalize them and leave phenotype character unchanged.OBJECTIVE: To establish immortalized precartilaginous stem cells (PSCs) from neonatal SD rats in vitro for the further related research about the differentiation mechanism and clinical application of precartilaginous stem cells.DESIGN,TIME AND SETTING: Single sample observation.The study was carried out in the Department of Orthopedics.Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from October 2005 to September 2006.MATERIALS: Neonatal SD rats,irrespective of gender,24-hour old,were used for prepare PSCs.METHODS: By using LipofectamineTM 2000,a gene transfection reagent,plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigene gene (SV40Tag) was transfected into the primary cultured PSCs isolated by immuniomagnetic beads coasted with the second antibody.Colonies were isolated by puromycin selection and expanded by many passages.MAIN OUTCOME MEASURES: Biological character of PSCs; plasmid identification; biological character of transfected cells and identification; RT-PCR; growth curve.RESULTS: Immunomagnetic beads separation system obtains PSCs,which was confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive PSCs.Double restriction enzyme was cut,electrophoresis confirmed pCMV was 3 kb,SV40T was 2.3 kb.A particular anti-puromycin cell clone was acquired,which was confirmed as FGFR-3 positive PSCs.The total RNA was isolated from the positive cell clones,and a 588 bp fragment,which was specific for the SV40T antigene gene,was amplified.The transfected cells were expanded to immortalized cell strain,named as immortalized precartilaginous stem cells (IPSCs).Thepopulation doubling time of IPSCs was (22.98±2.77) hours,no significant effect of subculture,freezing and recovering had been found.CONCLUSION: Precartilaginous stem cells could be isolated from neonatal SD rats,cultured in vitro,and immortalized through the transfection of pCMVSV40T/PUR.

Exploration and practice of PBL interactive teaching mode for overseas students / 中华医学教育探索杂志

PBL,problem based learning,is a kind of teaching model for resolving problems. Based on the present teaching situation of our university,my experience in NewYork University and my teaching experience for overseas students,this article elucidated the importance and necessity of PBL teaching model in overseas student education and put forward a suitable PBL teaching method.