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Objective:To investigate the risk factors of bone cement leakage and recompression of injured vertebrae after percutaneous kyphoplasty (PKP) for osteoporotic vertebral compression fracture (OVCF).Methods:A case-control study was performed to analyze the clinical data of 297 patients with single-segment OVCF who underwent PKP in First Affiliated Hospital of Soochow University from January 2017 to January 2021, including 67 males and 230 females; aged 60-92 years [(69.5±8.2)years]. According to the occurrence of bone cement leakage, the patients were divided into leakage group ( n=36) and no leakage group ( n=261). According to the occurrence of recompression of injured vertebrae, the patients were divided into recollapse group ( n=40) and no recollapse group ( n=257). The gender, age, fracture segment, type of fracture, fracture severity, cortical disruption, intravertebral cleft, preoperative and postoperative local kyphosis angle, correction value of local kyphosis angle, bone cement injection volume, bone cement distribution, and postoperative anti-osteoporosis treatment were recorded. Univariate analysis was used to analyze the correlation of those factors with bone cement leakage and recompression of injured vertebrae after PKP, followed by multivariate Logistic regression analysis to identify the independent risk factors. Results:Univariate analysis showed that fracture severity, cortical disruption and bone cement injection volume were related to bone cement leakage ( P<0.05 or 0.01). Gender, age, fracture segment, type of fracture, intravertebral cleft, preoperative and postoperative local kyphosis angle, correction value of local kyphosis angle, bone cement distribution, and postoperative anti-osteoporosis treatment were not related to bone cement leakage (all P>0.05). Univariate analysis showed that intravertebral cleft, bone cement distribution, and postoperative anti-osteoporosis treatment were associated with recompression of injured vertebrae (all P<0.01). Gender, age, fracture segment, type of fracture, fracture severity, cortical disruption, preoperative and postoperative local kyphosis angle, correction value of local kyphosis angle, and bone cement injection volume were not related to recompression of injured vertebrae (all P>0.05). Multivariate Logistic regression analysis showed that severe fracture ( OR=4.23, 95% CI 1.52-11.81, P<0.01), cortical disruption ( OR=3.29,95% CI 1.52-7.13, P<0.01), and bone cement injection volume >8 ml ( OR=2.31,95% CI 1.09-4.92, P<0.05) were significantly related to bone cement leakage. Multivariate Logistic regression analysis showed that intravertebral cleft ( OR=2.10, 95% CI 1.03-4.30, P<0.05), solid type of bone cement distribution ( OR=2.56, 95% CI 1.25-5.27, P<0.05) and no anti-osteoporosis treatment after operation ( OR=3.06, 95% CI 1.46-6.40, P<0.01) were significantly related to recompression of injured vertebrae. Conclusions:For OVCF patients, severe fracture, cortical disruption, and bone cement injection volume>8 ml are independent risk factors for bone cement leakage after PKP. Intravertebral cleft, solid type of bone cement distribution, and no anti-osteoporosis treatment after operation are independent risk factors for recompression of injured vertebrae after PKP.
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Objective To construct a human renal epithelial cell line HEK293T by CRISPR-Cas9-based site-directed knock-in of vascular endothelial growth factor 165 (VEGF165) gene, and avoid the off-target effect caused by lentivirus infection. Methods The VEGF165 expression vector with homologous arm (pUCm-T-VEGF165 plasmid) and the sgRNA expression vector [pSpCas9(BB)-2A-Puro-sgRNA plasmid] were designed and constructed based on the DNA sequence of the EZH2 gene, and then co-transfected into HEK293T cells. The expression of VEGF165 mRNA was detected by qPCR and the expressions of VEGF165 proteins were detected by Western Blot. Results The qPCR and Western Blot results showed that, comparing with the control, the pUCm-T-VEGF165 plasmid and pSpCas9(BB)-2A-Puro-sgRNA plasmid, the expression of the co-transfection plasmid were significantly increased, i.e. 3.42±0.30 vs. 1.02±0.21, 1.13±0.16 and 0.98±0.18 for the VEGF165 mRNA level (all P<0.01), and 1.13±0.16 vs. 1.02±0.06, 0.88±0.03 and 0.80±0.05 for the VEGF165 protein level (all P<0.01), respectively. Besides, the expression of EZH2 was significantly down-regulated, i.e. 0.14±0.06 vs. 1.08±0.11, 1.02±0.12 and 1.13±0.16 for the EZH2 mRNA level (all P<0.01), and 0.23±0.03 vs. 1.05±0.13, 0.91±0.04 and 0.81±0.06 for the EZH2 protein level (all P<0.01), respectively. This result showed that the VEGF165 was successfully inserted into the EZH2 genome, interfering the EZH2 expression. Conclusions VEGF165 gene can be successfully knocked into HEK293T cells by CRISPR/Cas9 system.
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Objective@#To evaluate the clinical efficacy of percutaneous kyphoplasty (PKP) in the treatment of stage Ⅲ Kümmell＇s disease with no neurological symptom.@*Methods@#From January 2009 to June 2018, 45 patients underwent PKP for stage Ⅲ Kümmell＇s disease with no neurological symptom at Department of Orthopaedics, The First Hospital Affiliated to Soochow University. They were 12 males and 33 females, aged from 61 to 85 years (average, 70.9 years). Their visual analog scale (VAS) scores for back pain, Oswestry disability indexes (ODI), heights of fractured vertebrae and kyphosis cob angles were recorded and compared between preoperation, one day, 3 months and final follow-up after operation.@*Results@#All the 45 patients were followed up for 12 to 48 months (average, 28.0 months). Their VAS scores (2.4±0.7, 2.2±0.7 and 2.3±0.6), ODI sores (34.7±6.8, 34.2±6.5 and 34.1±6.6) and cobb angles (15.7°±2.2°, 15.8°±2.2° and 15.9°±2.4°) at one day, 3 months and final follow-up after operation were significantly lower than those (8.2±1.1, 75.3±9.0 and 25.2°±3.9°) before operation (P<0.001). Their anterior height ratios of the fractured vertebra (54.0%±4.3%, 53.7%±4.2% and 53.6%±4.0%) and median height ratios of the fractured vertebra (56.8%±4.0%, 56.5%±3.9% and 56.6%±3.9%) at one day, 3 months and final follow-up after operation were significantly higher than those (25.8%±3.9% and 27.2%±3.1%) before operation (P<0.001). The rate of cement leakage was 13.3%(6/45). No patients had neurological symptoms after operation.@*Conclusion@#PKP is a minimally invasive, safe and effective treatment for stage Ⅲ Kümmell's disease with no neurological symptom, because it can relieve the symptoms of the patients, restore the height of the fractured vertebral body and reduce the local kyphosis cobb angle.
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Objective To evaluate the clinical efficacy of percutaneous kyphoplasty (PKP) in the treatment of stage Ⅲ Kümmell's disease with no neurological symptom.Methods From January 2009 to June 2018,45 patients underwent PKP for stage Ⅲ Ktimmell's disease with no neurological symptom at Department of Orthopaedics,The First Hospital Affiliated to Soochow University.They were 12 males and 33 females,aged from 61 to 85 years (average,70.9 years).Their visual analog scale (VAS) scores for back pain,Oswestry disability indexes (ODI),heights of fractured vertebrae and kyphosis cob angles were recorded and compared between preoperation,one day,3 months and final follow-up after operation.Results All the 45 patients were followed up for 12 to 48 months (average,28.0 months).Their VAS scores (2.4 ±0.7,2.2 ±0.7 and 2.3 ±0.6),ODI sores (34.7 ±6.8,34.2 ±6.5 and 34.1 ±6.6) and cobb angles (15.7°± 2.2°,15.8° ± 2.2° and 15.9° ± 2.4°) at one day,3 months and final follow-up after operation were significantly lower than those (8.2 ± 1.1,75.3 ±9.0 and 25.2°±3.9°) before operation (P ＜ 0.001).Their anterior height ratios of the fractured vertebra (54.0％ ±4.3％,53.7％ ±4.2％ and 53.6％ ±4.0％) and median height ratios of the fractured vertebra (56.8％ ± 4.0％,56.5％ ± 3.9％ and 56.6％ ± 3.9％) at one day,3 months and final follow-up after operation were significantly higher than those (25.8％ ± 3.9％ and 27.2％ ±3.1％) before operation (P ＜ 0.001).The rate of cement leakage was 13.3％ (6/45).No patients had neurological symptoms after operation.Conclusion PKP is a minimally invasive,safe and effective treatment for stage Ⅲ Kümmell's disease with no neurological symptom,because it can relieve the symptoms of the patients,restore the height of the fractured vertebral body and reduce the local kyphosis cobb angle.
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Objective To research the effect of alkylation of glycerol phosphate synthase (AGPS) in isoproterenol (ISO) induced rat cardiac hypertrophy. Methods The pathological cardiac hypertrophy rat model was constructed by ISO intraperitoneal injection. Twelve healthy Sprague-Dawley rats (120～150 g) were divided into ISO group and control group randomly. In the ISO group, rats were injected with ISO (3 mg/kg) per day for two consecutive weeks. In the control group, rats were injected with normal saline (3 mg/kg) per day for two consecutive weeks. Changes of left ventricular diastolic diameter, left ventricular posterior wall thickness, left ventricular ejection fraction, left ventricular short-axis shortening rate and left ventricular mass were detected by echocardiography. The cross-sectional area of myocardial cells in rats was measured by hematoxylin-eosin staining. The expression of hypertrophic factors [atrial natriuretic peptide (ANP), myosin light chain-2V (MLC-2V), α-myosin heavy chain (α-MHC)] and AGPS were detected by Western Blot and real-time quantitative PCR (qPCR). Results The results of echocardiography showed that the cardiac hypertrophy rat model was successfully constructed. The results of hematoxylin-eosin staining showed that the myocardial cross-sectional area in the ISO group was significantly larger than that of the control group. The Western Blot and qPCR results indicated that the relative expression of protein and mRNA of hypertrophic factor and AGPS in the ISO group were both up-regulated comparing with that of the control group, and the differences were statistical significance (all P<0.05). Conclusions The rat model of pathological cardiac hypertrophy with up-regulated AGPS expression was successfully constructed providing a theoretical basis for further study on the role of AGPS in pathogenesis of pathological cardiac hypertrophy.
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Objective@#To evaluate and compare the caries prevention effectiveness of a resin-based sealant and a glass ionomer sealant on permanent first molars in 7- to 9-years-old school children.@*Methods@#A randomized controlled, examiner-blinded, 5-year clinical trial was conducted in 19 primary schools in Dalian, Liaoning province. Totally 419 children (with 664 first molars) who had at least one sound permanent first molar with deep fissures or fissures with signs of non-cavitated lesions were recruited and stratified and allocated randomly into three parallel groups. A resin-based sealant was used in Group R (136 children with 219 first molars) and a glass ionomer sealant was used in Group G (130 children with 218 first molars). The third group (Group N, 153 children with 227 first molars) with none of the two sealants applied was served as blank control. Follow-up examinations were conducted at 0.5, 1, 2, and 5 years after sealing in order to assess the retention of the sealants and the incidence of caries occurrence. Chi-square test was adopted to compare the differences of caries incidence among the 3 groups. Generalized estimating equations (GEE) was used to analyze the effective factors of caries incidence.@*Results@#During the 5-year follow-up period, 13.4% (23/172) of the teeth in Group R, 22.5% (40/178) of the teeth in Group G and 34.5% (57/165) of the teeth in Group N were diagnosed as having developed cavitated lesions. And the differences in the incidence of caries among the three groups was statistically significant (P<0.05). According to GEE, the resin-based sealant and the glass ionomer sealant were found to be the protection factors and the non-cavitated lesion before sealing was found to be the risk factor. The resin-based sealant was found to be the protection factor to the glass ionomer sealant when the blank control group was removed.@*Conclusions@#Both resin-based sealant and glass ionomer sealant could effectively prevent the permanent first molars from the occurrence of carious lesions in 5 years. The resin-based sealant was superior to the glass ionomer sealant in caries prevention.
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Objective To construct rat lysine-specific histone demethylase 1 (LSD1) overexpression plasmids and LSD 1 demethylation fragment disfunction plasmids,and to evaluate their expression levels in HEK293T cells.Methods LSD1 fragments were amplified by PCR,and LSD1 demethylation disfunction fragments were amplified by overlap PCR.Rat-specific LSD1 overexpression plasmids and LSD1 demethylation disfunction plasmids were constructed and verified by agarose gel electrophoresis and sequencing.HEK293T cells were infected using the validated recombinant plasmids and blank vectors,and the stably expressed cells were selected by puromycin.The expression of LSD1 in the stably expressed HEK293T was detected by Western Blot and real-time quantitative PCR.Results The results of agarose gel electrophoresis and sequencing showed that LSD1 overexpression plasmids and LSD1 demethylation disfunction plasmids were successfully constructed.The Western Blot and real-time quantitative PCR results showed that compared with the blank group,the relative expression of LSD1 and mRNA in the LSD1 overexpression group and the LSD1 demethylation disfunction group were up-regulated,and the differences were statistically significant(all P＜0.01).Conclusions The constructed LSD1 overexpression plasmids and LSD1 demethylationi disfunction plasmids can achieve overexpression of LSD1 gene in HEK293T cells.This paper lays a foundation for further study of the relationship between LSD 1 gene and related diseases and demethylation of LSD 1.
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Objective To evaluate the clinical effects of minimally invasive percutaneous new C2transpedicular lag-screw fixation for management of Hangman's fractures.Methods From October 2015 to June 2017,25 patients who had Hang-man's fracture were operated with minimally invasive percutaneous new C2transpedicular lag-screw fixation assisted by infrared navigation,among them there were 18 male and 7 female,aged from 28 to 70 years[average(46.1±13.3)years].According to the Levine-Edwards classification:16 patients were TypeⅠ,6 patients were TypeⅡand 3 patients were TypeⅡA.The cause of the in-jury was road traffic accident in 20 patients and fall from height in 5 patients.19 patients were simple Hangman's fracture and oth-er associated lesions included odontoid fracture of TypeⅡ(2 patients),atlas fracture of TypeⅠ(2 patients),rupture of spleen(1 pa-tient),and rib fractures(1 patient).According to American Association for spinal cord injury,graded as spinal cord injury D in 3 cases and E in 22 cases.The clinical outcomes were evaluated by visual analog scale(VAS)scores and related indicators,such as operation time,the range of motion(ROM)of cervical spine and ratio of bony union were recorded.The pre-and post-operative VAS scores of neck pain were compared with repeated measures analysis of variance. Results The mean follow-up time was (12.5±6.0)months,ranging from 3 to 24 months.Satisfactory reduction was obtained in all cases(minimal translation≤2 mm with-out obvious C2,3angulation).The average operation time was(65.9±12.1)min.At first day,one month,three months and the last fol-low-up, VAS scores of neck pain decreased comparing with preoperational measurements. There was significant difference be-tween the pre-and post-operative VAS scores of neck pain(t=24.7,25.8,23.1,24.1,P<0.001).According to American Associa-tion for spinal cord injury,three cases restored from D to E.There was no spinal cord or vertebral artery injury during operation and there was no screw loosing or breakage and cervical malformation happened during follow-up period. Bony fusion was achieved in all of these cases and the range of neck rotation was stored normal at the last follow-up.The average time of Bony fu-sion was(3.4±0.8)months.Conclusion The primary clinical efficacies of minimally invasive percutaneous new C2transpedicu-lar lag-screw fixation for treatment of Hangman's fracture were satisfactory,which carries precision and safety,immediate stability and more reliable.So it could be considered as a new choice for management of Hangman's fractures.
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Objective To construct the Hut78 cell line with EZH2 gene knocked into by CRISPR/Cas9 system. Methods The EZH2 expression vector pMD-18T-EZH2 with homologous arm and the sgRNA expression vector pSpCas9 (BB)-2A-Puro-sgRNA, which could cut the double stranded genomic DNA, were constructed, and the two vectors were co-transfected into Hut78 cells. Then the expression of EZH2 mRNA was detected by qPCR, and the expressions of EZH2 and H3K27me3 proteins were detected by Western blot assay. Results The pMD-18T-EZH2 and pSpCas9(BB)-2A-Puro-sgRNA recombinant vectors were confirmed by DNA sequencing. When Hut78 cells were transfected with the two recombinant plasmid, qPCR results showed that the expression of EZH2 mRNA was significantly increased, and Western blot analysis showed that the expressions of EZH2 and H3K27me3 proteins were significantly increased. Conclusion EZH2 gene is successfully knocked into Hut78 cells by CRISPR/Cas9 system.
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Objective To investigate the effect of cancer-associated fibroblasts (CAFs)-derived chemokine ligands 7 (CCL7) on the proliferation and invasion of triple-negative breast cancer (TNBC) cells. Methods The mRNA expression level and protein level of CCL7 in CAFs and paracancerous fibroblasts were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western Blot respectively. To confirm the paracrine level of CCL7 in CAFs and paracancerous fibroblasts, the protein levels of CCL7 in the corresponding conditional medium were detected through enzyme-linked immunosorbent assay (ELISA). The effect of CCL7 on the proliferation and invasion of MDA-MB-231 (TNBC cell line) was investigated by MTS assay and Transwell assay, respectively. Results In comparison with paracancerous fibroblasts, the mRNA expression level and protein level of CCL7 in CAFs were significantly increased (both P<0.01). There was an obviously increase of paracrine level of CCL7 in CAFs-conditional medium (P<0.01). The MTS assay and Transwell assay results indicated that CCL7 was more able to promote the proliferation and invasion of MDA-MB-231. Conclusion CAFs in the TNBC stroma can produce more chemokine CCL7, and CCL7 can promote the proliferation and invasion of TNBC cells
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<p><b>OBJECTIVE</b>To evaluate the effectiveness of resin-based sealants and glass ionomer sealants on caries prevention in the first permanent molars.</p><p><b>METHODS</b>A randomized controlled, examiner-blinded, 2-year clinical trial with parallel groups was conducted. A total of 419 children (664 first molars) who had at least one sound first permanent molar with deep fissures or fissures with signs of non-cavitated lesions were recruited. They were randomly allocated into three groups.Helioseal F, a light-cured resin-based sealants was used in group R (136 children with 219 first molars). FUJI VII, a light-cured glass ionomer sealants was used in group G (130 children with 218 first molars) and group N (153 children with 227 first molars) was blank control.Follow-up examinations were conducted at 6, 12, and 24 month after sealing to examine the sealants retention and caries incidence. Chi-square test was used to compare the differences of caries incidence among the 3 groups.Generalized estimating equations (GEE) was used to analyze the effective factors of caries incidence.</p><p><b>RESULTS</b>During the 2-year follow-up period, 3.0% (6/197) of the teeth in group R, 6.7% (13/193) of the teeth in group G, and 14.7% (29/197) of the teeth in group N were diagnosed as having developed a cavitated lesion. The difference in the incidence of caries among the three groups was significant(P < 0.05). No significant difference in caries incidence was found between group R and group G (P > 0.05). The tooth-level preventive fractions were 79.6% for resin-based sealants and 54.4% for glass ionomer sealants, respectively.</p><p><b>CONCLUSIONS</b>Both resin-based sealants and glass ionomer sealants can effectively prevent the incidence of caries in the first permanent molars. The two sealants have similar caries prevention effect.</p>
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Child , Humans , Acrylic Resins , Composite Resins , Dental Caries , Glass Ionomer Cements , Molar , Pit and Fissure Sealants , Silicon Dioxide , Single-Blind MethodABSTRACT
<p><b>OBJECTIVE</b>This study aims to determine the fissure morphology, prevalence, and severity of caries in the first permanent molars of 7-year-old to 9-year-old children in the Dalian development area. The study also aims to investigate the relationship between fissure morphology and noncavitated lesions.</p><p><b>METHODS</b>A multi-stage, cluster sampling method was used. Three schools were randomly selected from 20 elementary schools in the Dalian development area. All of the children in the selected schools completed clinical oral examinations. A diagnosis criterion of the International Caries Detection and Assessment System was used to assess the cavitated and noncavitated dental caries in the first permanent molars. Fissure depth and fissure stain were also assessed.</p><p><b>RESULTS</b>The total size of the study sample was 1 160. The prevalence of cavitated caries was 19.1%, and the mean D3-6FT score was 0.31. No significant differences were found between urban and rural subjects, between men and women, and among different age groups. The prevalence and D1-2FT score of noncavitated caries were 42.1% and 0.86, respectively. The D1-2FT score for rural children was significantly higher than that for urban children, and the D1-2FT score for girls were higher than that for boys. No statistical differences were found among different age groups. The prevalence of deep fissures in the first permanent molars was 44.7%. The prevalence of noncavitated caries increased with increasing fissure depth.</p><p><b>CONCLUSION</b>The prevalences of noncavitated caries and deep fissure in the first permanent molar of 7-year-old to 9-year-old children in Dalian were high. Fissure morphology was found to be related to noncavitated caries.</p>
Subject(s)
Child , Female , Humans , Male , China , DMF Index , Dental Caries , Molar , PrevalenceABSTRACT
Objective To establish a rapid and accurate analysis method for food-derived ACE inhibitory peptides activity in vitro.Methods Reaction time of ACE and substrate was by measuring the hippuric acid liberated in the ACE reaction mixture at regular intervals;An optimal RP-HPLC method to measure food-derived ACE inhibitory peptides activity in vitro was set up.The hippuric acid from ACE reaction mixture(sea cucumber peptides were regarded as ACE inhibitor) was estimated by Zorbax SB-C_(18) analytical column with acetonitrile and ultrapure water as mobile phase.Results The reaction time of ACE with substrate was determined at sixty minutes;The elution was carried out with the ratio of acetonitrile to ultrapure water was 1:1(0.1%TFA) at a flow rate of 0.4 mL?min~(-1).The ahsorbance of the eluent was monitored at 228 nm,and column temperature was 25℃.The relationship between hippuric acid concentration and peak area exhibited a good linearity in the concentration ranges of 0～200?g?mL~(-1) and 200～800?g?mL~(-1).The RP-HPLC method was further validated by captopril,the oyster hydrolysate and the anchovy hydrolysate.Conclusion The method has been proved to be convenient,accurate and suitable for the analysis of foodderived ACE inhibitory peptides activity in vitro.