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【Objective】 To explore the causal association between interleukin (IL) level and constipation by using two-sample Mendelian randomization. 【Methods】 Analyses were performed based on the data from gene-wide association studies (GWAS). Both interleukin and constipation data were obtained from European populations. IL as an exposure variable was obtained from two GWAS data sets: ⅰ. from a genetic map of the human plasma proteome containing 3 301 samples; ⅱ. from a GWAS data set on 90 circulating proteins, containing 30 931 samples. Constipation as an outcome variable was obtained from two GWAS data sets: ⅰ. from Finngene, containing 26919 cases and 282235 controls; ⅱ. from UKBiobank, containing a total of 3 328 cases and 459682 controls. Single nucleotide polymorphisms strongly associated with exposure variables were used as instrumental variables, with inverse variance weighted (IVW) as the main analysis method, MR-egger regression and weighted median method as supplementary evidence for IVW results, and horizontal pleiotropy and heterogeneity were tested to ensure the stability of the results. 【Results】 In both of the two different outcome variables GWAS data, IVW analysis results showed that decreased level of IL-17 receptor C was associated with an increased risk of constipation, with ORs of 0.956 (95% CI: 0.916-0.997, P=0.036‖Finngene) and 0.998 (95% CI: 0.997-0.999, P=0.040‖ukb). Increased level of IL-18 was associated with an increased risk of constipation, with ORs of 1.055 (95% CI: 1.008-1.104, P=0.022‖Finngene) and 1.001 (95% CI: 1.000-1.002, P=0.044‖ukb); while in the Finngene data, the IVW results also suggested that increased levels of IL-2 receptor alpha subunit α and decreased levels of IL-10 and IL-17 were associated with an increased risk of constipation, with ORs of 1.054 (95% CI: 1.001-1.110, P=0.049), 0.945 (95% CI: 0.896-0.996, P=0.035) and 0.934 (95% CI: 0.896-0.997, P=0.040). 【Conclusion】 IL-17 receptor C, IL-18, IL-2 receptor alpha subunit α, IL-10, and IL-17 were causally associated with the risk of constipation.
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【Objective】 To investigate the effects of biomimetic bone trabecular with the same porosity and pore size and regular porous structure on the adhesion, proliferation, and differentiation of osteoblasts, so as to provide theoretical basis for the improvement of osseointegration performance of titanium alloy implants. 【Methods】 The biomimetic bone trabecular and regular porous structures with the same porosity and pore size were generated by computer-aided software, and then processed into disc-shaped Ti6Al4V scaffolds with a diameter of 10 mm and a height of 3 mm by selective laser melting technology. MC3T3-E1 cells, the precursor cells of mouse osteoblasts in the logarithmic growth phase, were seeded on two kinds of scaffolds and divided into biomimetic bone trabecular group and regular porous structure group. After 3 hours of culture, acridine orange staining and phalloidin /DAPI staining were used to evaluate the number of cell adhesion. After 3 days of culture, the scaffolds were examined by scanning electron microscopy to evaluate the adhesion state of cells. After 1, 3, and 5 days of culture, the scaffolds were taken for CCK8 detection to observe the proliferation of cells. After 7 and 14 days of differentiation, alkaline phosphatase (ALP) activity was detected. After 14 days of differentiation, the expressions of osteogenesis-related genes (ALP, OCN, RUNX2) were detected by RT-PCR. After 30 days of differentiation, the scaffolds were stained with alizarin red and 100 g/L cetylpyridinium chloride was used to dissolve mineralized nodules. Calcium salt deposition was qualitatively and quantitatively detected to evaluate cell differentiation. 【Results】 The results of acridine orange and phalloidin /DAPI staining showed that the biomimetic trabecular Ti6Al4V scaffold adhered to more MC3T3-E1 cells than the regular porous structure, and the cytoskeleton of the former scaffold was more densely distributed. The results of scanning electron microscopy showed that the pseudopodia of MC3T3-E1 cells on the biomimetic bone trabecular Ti6Al4V scaffold were longer and the extension state was better than that of the regular porous structure. CCK8 test showed that the proliferation of MC3T3-E1 cells on the biomimetic trabecular bone titanium alloy scaffold was significantly higher than that on the regular porous structure on the 3rd and 5th day, and the difference gradually increased with the increase of time, with statistical significance (P<0.05). The results of cell differentiation test showed that ALP activity on the bionic trabecular scaffold was higher than that on the regular porous structure (P<0.05). The expressions of osteogenic genes (ALP, OCN, RUNX2) in MC3T3-E1 cells on the biomimetic bone trabecular titanium alloy scaffold were significantly higher than those on the regular porous structure (P<0.05). After 30 days of induction, the amount of calcium salt deposited in the bionic trabecular titanium alloy scaffold was significantly larger than that in the regular porous structure (P<0.05). 【Conclusion】 The biomimetic bone trabecular with a porosity of 65% and an equivalent pore size of 600 μm is more conducive to the adhesion, proliferation and differentiation of mouse osteoblast precursor cells MC3T3-E1 on the titanium alloy scaffold than the regular porous structure with the same porosity and pore size. It is theoretically more conducive to improving the osseointegration performance of titanium alloy implants.
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【Objective】 To investigate the clinical effects of treatment of single-segment lumbar tuberculosis by oblique lateral interbody fusion with autologous iliac bone and percutaneous pedicle screw fixation. 【Methods】 We collected the clinical data of 47 patients with lumbar tuberculosis treated in The First Affiliated Hospital of Xi’an Jiaotong University from March 2017 to January 2020. Among them， 22 patients underwent oblique lateral interbody fusion with autologous iliac bone and percutaneous pedicle screw fixation （minimally invasive group） and 25 patients underwent open surgery combined anterior-debridement and posterior-fixation （control group）. The related data were collected， including gender， sex， body mass index （BMI）， systemic symptoms of tuberculosis， operation duration， intraoperative bleeding， postoperative drainage， hospital stay， complications， visual analogue score （VAS）， erythrocyte sedimentation rate （ESR）， and Oswestry disability index （ODI）. 【Results】 Baseline clinical characteristics did not significantly differ between the two groups （P>0.05）. Compared with control group， the minimally invasive group had shorter operation duration ［（188.64±18.59） min vs. （201.60±22.67） min］， less intraoperative blood loss ［（118.64±22.95） mL vs. （553.60±100.54） mL］， less postoperative drainage ［（134.55±36.48） mL vs. （291.20±61.53） mL］， and shorter hospitalization time ［（12.86±2.17） d vs. （15.80±3.03） d］ （all P0.05）. Compared with the preoperative ones， ESR， VAS score and ODI score significantly decreased and Cobb angle significantly increased in both groups （all P0.05）. 【Conclusion】 Both minimally invasive technique and open surgery can achieve excellent clinical results， but the minimally invasive technique can reduce the surgical trauma and shorten the hospitalization time.
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Objective To investigate the etiologic roles of apoptosis-associated genes,environmental factors and their interactions in lumbar disc herniation (LDH).Methods A case-control trial was conducted.We recruited 128 outpatients with LDH as case group and 132 normal people matched by age and gender as control group.Peripheral venous blood samples were collected and DNA was extracted from leukocytes.By using a modified Brucker Autoflex MALDI-TOF mass spectrometer,we analyzed 3 genes with 9 polymorphic sites,namely,Fas-1377G/A rs2234767,Fas-670G/A rs1800682,Fas rs2147420,Fas rs2296603,Fas rs7901 656,Fas rs1 57101 9,FasL-844C/T rs7631 10,CASP-9-1263A > G rs4645978,and CASP-9-712C > T rs4645981.The correlations between polymorphism of Fas,FasL and CASP-9 genes and the risk of LDH were evaluated by non-conditional Logistic regression model.Multiple Logistic regression model was performed to assess the interaction between apoptosis-associated genes and environment factors,such as lumbar vertebral loads,bed type,spare-time exercises and spare-time activities. Results There were preferable balances in case and control groups in age and gender without significant differences.However,the two groups differed significantly (P G (rs4645978),and FasL-844C/T TT and CASP-9-1263A>G GG genotypes might be the high risk genotypes of LDH.The gene-environment interaction analysis revealed that super-multiplicative and sub-multiplicative interactions respectively between FasL-844TT genotype and lumbar vertebral loads (3-4 level),and between CASP-9-rs4645978 GG and lumbar vertebral loads (3-4 level).Conclusion FasL,CASP-9 genes and lumbar vertebral loads and their interactions play important roles in the pathogenesis of LDH.It suggests that the risk of LDH may be codetermined by environmental factors and inherited susceptibility genes,and that the mechanisms of interactions vary in different genotypes and the same or different environmental factors.
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This study examined the correlation between osteoporosis and the degeneration of intervertebral discs. Sprague-Dawley rats were maintained up to 22 or 28 months. The femoral bone, tibial bone and lumbar vertebra were histologically studied and the expression of collagen type II and X in intervertebral discs was immunohistochemiscally determined. Several indices for the degeneration of intervertebral discs and osteoporosis and the correlation among them were then analyzed. Close correlations were found among the indices for the degeneration of intervertebral discs, including the relative area of the vascular bud, the ratio of the uncalcified and the calcified layers, the expression of collagen type II and X. The correlation with collagen type X was negative. There existed positive correlations among the indices for osteoporosis, including the thickness ratio of cortical bone, the relative area of bone trabecula, the density of femoral and vertebral body bones, and the maximum stress and strain on bone. Analysis on the relationship of osteoporosis and the disease on disc showed that the indices of osteoporosis were negatively correlated with the indices of the degeneration of intervertebral discs but the expression of collagen type X was positively correlated, with the density of vertebral body bones having the strongest dependence on collagen type X. The maximum stress and strain bore no correlation with the degeneration of intervertebral discs. These results suggest that osteoporosis was negatively correlated with the degeneration of intervertebral discs.
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Objective To establish an animal model of crush syndrome in rabbits and investigate the preventive effect of negative pressure drainage.Methods Twenty-four New Zealand rabbits were randomly divided into four groups:control group,traditional treatment group,negative-pressure treatment group and mixed treatment group,with six rabbits per group.Blood urea nitrogen (BUN),creatinine (Cr),K,creatine kinase (CK) and myoglobin (Myo) were detected before crush and at 6,12,24,48,72 hours after decompression.Edema and analgesia of extremities were determined and changes of kidney and muscular tissue were observed by light microscope.Results As time went on,K,Myo,BUN and CR had unobvious rise in each treatment group,without the presence of renal failure.The control group showed marvelous increases of K,Myo,BUN and Cr at 6 hours after decompression,from (4.61 ±0.98)mmol/L,(4.22±0.93) ng/ml,(7.76±1.40) mmol/L,(101.32±9.35)μmol/Lto (7.88±1.95)mmol/L,(11.34 ±3.86) ng/ml,(15.91 ± 1.76) mmol/L,(258.32 ±91.40) μmol/L separately,with renal failure and significant differences in comparison with other treatment group (P ＜0.05).The level of tissue edema and degree of analgesia in negative-pressure treatment group and mixed treatment group were lower than those in the traditional treatment group.Renal damage in all the treatment groups was milder than that in the control group by light microscopic observation,while there was no significant difference of muscle injury among groups.Conclusion Negative pressure drainage may prevent crush syndrome.
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Objective By exploring the effects of recombinant human bone morphogenetic protein (rhBMP)2 and rhBMP13 on chondrocytes proteoglycan production and phenotype expression to establish the theoritical mechanisms for the treatment of disc degeneration with chondrocytes transplantation plus BMPs.Methods The dose-dependent effects of rhBMP2 and rhBMP13 on PG protein synthesis and gene expression were detected under different concentrations (0,25,125,and 625 ng/ml).The sulfated-glycosaminoglycan (s-GAG) in the culture media and the pericellular matrix was measured with a 1,9-dimethyl-methylene blue (DMMB) colorimetric assay.Reverse transcriptase polymerase chain reaction (RT-PCR) was performed respectively to quantify the relative abundance of aggrecan Mrna.Cell proliferation was examined by Hoechst Dye assay.Results All rhBMP2 and rhBMP13 in different concentrations could significantly increase s-GAG synthesis and gene expression in chondrocytes (P＜0.05).And at the same concentration.rhBMP2 was more potent than rhBMP13 on s-GAG synthesis.Hoechst Dye assay showed neither rhBMP2 nor rhBMP13 had significant effect on cell proliferation.Conclusion rhBMP2 and rhBMP13 are able to upregulate s-GAG synthesis,in addition,rhBMP2 is more potent than rhBMP13 on aggrecan gene expression regulation,but rhBMP2 and rhBMP13 do not have significant effect on chondrocyte proliferation.
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Objective To develop retrovirus-mediated transfer of the fusion gene encoding EGFP-BMP2 in mesenchymal stem cells. Methods Mesenchymal stem cells from New Zealand white rabbits were transduced with retroviral pLEGFP-BMP2 vector by the optimized retroviral transduction protocol. Fluorescent microscopy's examination was to evaluate the results of the transduction, flow cytometer's analysis was to evaluate the transduction efficiency and the Fluorescence-activated cell sorting method was to sort the transduced cells. Bioactivity test from C2C12K4 cells was to show the expression and bio-activity of the fusion gene. Results Fluorescent microscopy showed the success of the transduction. By flow cytometer's analysis, the mean efficiency of the transduction with EGFP was (42.8±6.1)% SD. Transduced cells were sorted efficiently by the fluorescence-activated cell sorting method and after sorting, almost of those showed the expression of BMP2. Fluorescently and strongly bioactivity test for C2C12K4 cells demonstrated that fluorescent materials were located the surface of cells and the activity of luciferase increased compared with the control. Analysis of long-term expression showed there was no difference between 2 week-time point and 3 month-time point of culture post-sorting. Conclusion Mesenchymal stem cells can be transduced efficiently by retrovirus-mediated transfer of the fusion gene encoding EGFP-BMP2, the highly pure transduced cells are obtained by the fluorescence-activated cell sorting technique, the expressed chimeric protein embraced the double bioactivity of EGFP and BMP2, and moreover, the expression had not attenuated over time.
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Objective: To develop retrovirus-mediated transfer of the fusion gene encoding EGFP-BMP2 in mesenchymal stem cells. Methods: Mesenchymal stem cells from New Zealand white rabbits were transduced with retroviral pLEGFP-BMP2 vector by the optimized retroviral transduction protocol. Fluorescent microscopy's examination was to evaluate the results of the transduction, flow cytometers analysis was to evaluate the transduction efficiency and the Fluorescence-activated cell sorting method was to sort the transduced cells. Bioactivity test from C2C12K4 cells was to show the expression and bio-activity of the fusion gene. Results: Fluorescent microscopy showed the success of the transduction. By flow cytometer's analysis, the mean efficiency of the transduction with EGFP was (42. 8 ± 6. 1)% SD. Transduced cells were sorted efficiently by the fluorescence-activated cell sorting method and after sorting, almost of those showed the expression of BMP2. Fluorescently and strongly bioactivity test for C2C12 K4 cells demonstrated that fluorescent materials were located the surface of cells and the activity of luciferase increased compared with the control. Analysis of long-term expression showed there was no difference between 2 week-time point and 3 month-time point of culture post-sorting. Conclusion: Mesenchymal stem cells can be transduced efficiently by retrovirus-mediated transfer of the fusion gene encoding EGFP-BMP2, the highly pure transduced cells are obtained by the fluorescence-activated cell sorting technique, the expressed chimeric protein embraced the double bioactivity of EGFP and BMP2, and moreover, the expression had not attenuated over time.
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Objective To investigate the apoptosis of articular chondrocyte and the expression of Bcl-2, Bax, Fas and iNos in articular cartilage with Kashin-Beck disease(KBD) in order to understand the pathogenesis of chondronecrosis in KBD. Methods The collected samples of human articular cartilage were divided into two groups: control group (15 samples from 15 cases), KBD group (15 samples from 15 cases). KBD patients were diagnosed by "Pathological Criteria to Diagnose KBD in China". Chondrocyte apoptosis was detected by TUNEL staining, and the Bcl-2, Bax, Fas and iNos positive articular chondrocytes were stained by the B-SA of immunohistochemistry. Articular cartilage was classified three zones and the positive rate were counted by light microscope for cytoplasimic staining by polyclonal antibodies of Bcl-2, Bax, Fas and iNos and apoptotic chondrocytes by TUNEL. Results ① The percentage of positive apoptotic chondrocytes stained by TUNEL in the middle zone of articular cartilage from the KBD-children group(33.60±2.71%) was higher than that of the control (1.33±0.41% t=11.59, g=28, P<0.01). ②The percentage of chondrocytes staining for Bcl-2, Bax, Fas and iNos among the upper and the middle zone in KBD group were significantly higher than that of the control (t=11.75-18.65, g=14, P<0.01); the remarkable difference in the expression of Bcl-2, Bax, Fas and iNos among the upper, the middle and the deep zones was also seen in KBD articular cartilage (F=73.49-114.42, g=42, P<0.01), and staining for Bcl-2, Bax, Fas and iNos in KBD children was prominent in the upper zone(41.93±12.26%, 45.60±15.78%, 53.60±16.49%, 45.47±14.02%) and the middle zone(14.93±3.50%, 13.87±4.32%, 23.27±4.83%, 21.67±6.82%)of articular cartilage, respectively. Conclusion The chondrocyte apoptosis and the present of Bcl-2, Bax, Fas and iNos positive chondrocytes in articular cartilage of children with KBD were significantly higher than that of the control.
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<p><b>OBJECTIVE</b>To observe clinical efficacy of intra-articular injection of Sodium hyaluronate (SH) accompanied with external application of Sanhua ointment (SHO) for knee osteoarthritis.</p><p><b>METHODS</b>One hundred and twelve patients with osteoarthritis were randomly divided into two groups, Group I was treated with SH and Group II was treated with SH plus SHO. The entire condition of knee joint in the two groups were compared before and after treatment according to Lysholm's function scoring.</p><p><b>RESULTS</b>The functional score in the two groups at the 2nd week, 5th week, 3rd month and end of 1st year of the treatment course were significantly higher than that before treatment (P < 0.05), and the scores continuously increased within the first 3 months of treatment. Comparison of scores between the two groups showed that scores in Group II was significantly higher than those in Group I at the corresponding period. The total effective rate after 1 year treatment in Group lI was higher than that in Group II (81.5% vs 96.6%, P < 0.05). In Group II , the initial time of symptom improving was earlier and the adverse effect disappeared more quickly than those in Group I markedly.</p><p><b>CONCLUSION</b>The efficacy of combined therapy of intra-articular injection of SH and SHO for knee osteoarthritis was superior to that of intra-articular injection of SH only.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Administration, Topical , Drug Therapy, Combination , Drugs, Chinese Herbal , Hyaluronic Acid , Injections, Intra-Articular , Osteoarthritis, Knee , Drug Therapy , PhytotherapyABSTRACT
Objective To study the correlation between osteoporosis and degeneration of intervertebral disc in the aging rat.Methods Fifty healthy SD rats were fed for twenty-two months in order to build the model of aging rats,the histology of femoral bone,tibial bone and lumbal vertebra was used to observe,then we evaluated the indexes which reflect the degeneration of intervertebral disc including the quantity of collagen type Ⅱ,Ⅹ of intervertebral disc by the immunohistochemistry methods,the relative area of vascular bud,the thickness of calcified layer and un-calcified layer on cartilage endplate and the indexes which reflect the osteoporosis including the thickness ratio of cortical bone,the relative area of bone trabecula,the bone density,the max stress and straining of bone.Results There was moderate or strong negative correlation among the indexes of the degeneration of intervertebral disc including the relative area of vascular bud,the ratio of the un-calcified and the calcified layer,the quantitation of collagen type Ⅱand the indexes of osteoporosis including the thickness ratio of cortical bone,the relative area of bone trabecula,the density of femoral bone and vertebral body bone,the max stress and straining of bone.However,there was moderate or strong positive correlation with the quantitation of collagen type Ⅹ,of which the bone density of vertebral body had the strongest dependablity with collagen type Ⅹ.And the indexes of bone biomechanics including the max stress and straining had no correlation with the indexes of degeneration of intervertebral disc.Conclusion Osteoporosis has negative correlation with the degeneration of intervertebral disc,which is not distributed to the environment factors;but the indexes of bone biomechanics including the max stress and straining have no correlation with the indexes of degeneration of intervertebral disc.
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@#ObjectiveTo investigate the effect of operation treating spinal multi-level tuberculosis.Methods45 patients with multi-level spine tuberculosis were treated with debridement completely and anterior or lateral-anterior intervertebrae autograft. Of them, 5 patients added to anterior instrumentation.Results45 patients had been followed up for 12 to 40 months.The back pain of 40 cases were relieved within 5 weeks after operation, erythrocyte sedimentation rate decreased 3 weeks after operation. 38 patients who involved in kyphosis decreased their kyphosis mostly and only one patient lost 3 degrees in follow-up.15 patients who involved in neurological deficits improved one or two grades (Frankel). Grafts fused in 44 patients and there were no recurrent in follow-up.Conclusions Operative treatment is efficacious to multi-level body spine tuberculosis.
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Objective To explore the preventive effect of the supplemental dietary boron on bone damage of rats with excess fluoride (EF) ingestion. Methods Twenty-four Sprague-Dawley rats aged 4-5 weeks old were divided into the control (C group, treated with distilled water and the elementary dietary), the excessive fluoride dose group (EF group, treated with distilled water with 100mg/L F-from 221.0g NaF per liter and the elementary dietary) and the boron prevention group (P group, treated with 100mg/L F-distilled water and the supplemental boron dietary). Three months after the experiment, fluorine, boron contents and AKP activities in serum, total RNA and fluorine contents in bone, vertical and transverse diameter and dry weight of tibias of rats were assayed, and bone biomechanics of femur and bone mineral density (BMD) were determined. Results Compared with those in the C group, fluorine contents in serum and bone, AKP activities in serum of rats in the EF group significantly increased; total RNA in bone and BMD of the bone of whole body and tibia decreased, vertical and transverse diameter of tibias was shortened, dry weight was decreased; peak load of femur increased but maximum deformation decreased. Compared with those in EF group, fluorine contents in serum and bone, AKP activities in serum of rats in the P group obviously decreased; total RNA in bone and BMD of whole body, lumbar vertebrae and tibias notably increased, vertical and transverse diameters and dry weight of tibia were enhanced, peak load of femur was obviously raised. Boron contents in serums of rats in three groups had no significant differences. Conclusion Bone damage obviously occurred in rats with EF intake, and the supplemental dietary boron had a preventive effect on these changes.
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Objective To determine the potential value of imaging for spinal tuberculosis. Methods 180 patients proved as spinal tuberculosis by operation or clinical follow who underwent X-ray film, CT and MRI were reviewed. They were classified A, B or C in term of imaging and clinical symptom. A was the normal of X- ray film and positive of CT or MRI. B was positive of X-ray film, CT and MRI. C was with the neurological symptoms. Results 40 patients ( 40/180 ) were categorized as A . They had short duration (
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Objective To investigate the effects of gene transfection with human growth/differentiation factor 5(hGDF5)on the differentiation and proliferation of rabbit bone marrow mesenchymal stem cells (BMSCs).Methods BMSCs were obtained from adult New zealand rabbits and purified by gradient centrifuge.Exogenous recombinant human GDF5 was transfected into BMSCs with liposome method.Then hGDF5 expression at mRNA and protein level was measured separately by reverse transcription-polymerase reaction and indirect immunofluorescence method.Activity of alkaline phosphates(ALP),expression of TypeⅡcollagen(ColⅡ),proteoglycan and growth of the cells were all measured by biological methods to evaluate the effects of hGDF5 gene transfer on the differentiation and proliferation of rabbit BMSCs.Results After hGDF5 gene transfection,BMSCs expressed hGDF5 mRNA and protein,and compared with the control groups,expression of proteoglycan and ColⅡ increased significantly,but no significant difference appeared in ALP activity and cell proliferation.Conclusion Gene transfer with hGDF5 is an effective way to enhance the expression of GDF5 at mRNA and protein.The expression of heterogenetic hGDF5 gene can induce BMSCs' differentiation to chondrogenic cells.But the gene transfection has no obvious effects on the proliferation and ALP activity of BMSCs.
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Objective To establish the animal model of cervical pain induced by formalin injection and explore the effect of cervical pain on cervical vertigo.Methods Formalin of 40 mL/L was injected near the nerve root of left side of neck to establish the model of cervical pain in rabbits.A total of 30 rabbits were divided randomly into two groups.Formalin-induced pain group(15 rabbits): indexes related to the vertebral artery were observed respectively before 2 mL of formalin injection and after it at 5,10 and 30 minutes.Lidocaine-blocking pain group(15 rabbits): pain induced by formalin was blocked by lidocaine of 10 g/L after 15-minute drug administration and the same indexes were observed at the same time and position of the neck.Results In formalin-induced pain group,vertebral artery systolic velocity,diastolic velocity and pulsatility indexes were decreased whereas resistance index was increased(P