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AIM: To investigate the effect of inhibiting Ca2+/calmodulin-dependent protein kinase Ⅱ(CAMKⅡ)expression in adult retinal pigment epithelial cell line-19(ARPE-19)cells on the migration, invasion, and tube formation of human umbilical vein endothelial cells(HUVECs)in a non-contact co-culture system.METHODS: RNA sequencing was performed on ARPE-19 cells overexpressing CAMKⅡ-δ, and bioinformatics was used to analyze the biological functions of the differentially expressed genes. Transwell inserts was used to construct a non-contact co-culture system of ARPE-19 and HUVECs. The experimental groups included: blank group: only HUVECs were inoculated without ARPE-19 cells; control group: ARPE-19 and HUVECs cells were co-cultured with complete medium; AIP group(CAMKⅡ inhibition group): ARPE-19 cells in AIP(160 nmol/L)were co-cultured with HUVECs in complete medium. The migration, invasion and tube formation abilities of HUVECs were detected. The protein expression levels of CAMKⅡ/AMPK/mTOR/VEGFA were detected by Western blotting.RESULTS:Bioinformatics analysis found that the differentially expressed genes could affect biological processes such as cell growth and death and cell movement. The scratch test and transwell migration test showed that the relative mobility of HUVECs in the AIP group was significantly lower than that in the control group(all P<0.05). However, the invasion and tube formation assay showed that the relative invasion rate and tube formation rate of the AIP group were not significantly different from those of the control group(both P>0.05). Western blotting results showed that the expression levels of CAMKⅡ, P-mTOR, and VEGFA proteins in the AIP group were significantly lower than those in the control group, while the expression level of the P-AMPK protein was significantly higher than that in the control group(all P<0.05).CONCLUSION:In the non-contact co-culture system, inhibition of CAMKⅡ expression in ARPE-19 cells significantly reduced the migration ability of HUVECs, but it cannot change the invasion and tube formation ability, which may be achieved by AMPK/mTOR/VEGFA.
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Objective To evaluate the clinical outcomes of minimally invasive internal fixation for femoral intertrochanteric fracture in osteoporotic elderly patients. Methods A retrospective study was conducted in 112 patients using interan nail (IN) or trochanteric antegrade nail (TAN) for the management of intertrochanteric femoral fracture from January 2009 to September 2015 in our hospital. According to AO classification, there were 34 cases of type 31-A.1, and 61 cases of type 31-A.2, 17 cases of type 31-A.3. Clinical and radiological follow-up were available. Surgical and fluoroscopic time, length of hospital stay, blood loss, complications and hip functions were compared between two groups. Results A total of 78 patients meeting the criteria were evaluated at a mean follow-up of 15 months (range, 3-27 months). The IN was used in 41 patients and the TAN in 37 patients. Operative time, fluoroscopy time and blood loss showed significant difference between the IN group and TAN group (respectively, 58.9±6.9 vs. 75.6±5.9min; 2.70±0.47 vs. 4.40±0.47min; 107.6±6.7 vs. 127.8±6.8ml, P0.05). Conclusions For minimally invasive treatment of unstable femoral intertrochanteric fractures, use of either the IN or TAN is clinically effective. However, IN presents more advantages (e.g., easy operation, reliable fixation, less bleeding, better clinical outcomes, and less complications). The use of IN is a suitable option for the treatment of unstable intertrochanteric fractures in osteoporotic elderly patients.
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Objective To evaluate the effect of TGF-β3 on rabbit nucleus pulposus( NP) cells cultured in three-dimensional polylactic-co-glycolic acid (PLGA)scaffold in vitro.Methods PLGA scaffolds were fabricated by particulate leaching method and soaked in rabbit NP cells suspension(1 × 106/scaffold).PLGA-seeded NP cells were devided into 4 groups: 100 ng/mL TGF-β3/PLGA,500 ng/mL TGF-β3/PLGA,1 μg/mL TGF-β3/PLGA, PLGA control group.Cell proliferation activity was measured using MTT assay.The glycosaminoglycan ( GAG ) analysis were performed by 1, 9-dimethylmethylene blue(DMMB) assay.mRNA expression was measured by quantitive PCR at each time point.Histological observation was performed to elucidate the morphological changes of NP cells in PLGA effected by TGF-β3.Results Higher cellular proliferation activity, GAG production,Collagen type II, Aggrean expression were observed in TGF-β3 /PLGA-seeded NP cells compared with PLGA control group on day-7,day-14,day-21(P<0.05). Higher dose of TGF-β3 exhibited intense cellular proliferation activity and peri-cellular matrix by increasing trend(P<0.05).Histological observation showed TGF-β3/PLGA developed more significant disc cells cluster than PLGA groups on day-21.Conclusion The 3D porous PLGA scaffold-seeded cells using TGF-β3 can promotes cell proliferation, and prompt extracellular matrix(ECM) production.It is a potential biotherapy for the treatment of disc degeneration.
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Objective To compare the clinical and radiographic outcomes of 3 kinds of lumbar interbody fusion (ALIF,PLIF,TLIF) for adult low-grade isthmic spondylolisthesis at L4.5 and L5S1 levels.Methods Data of seventy patients with lowgrade isthmic spondylolisthesis who had undergone different lumbar interbody fusion (LIF) procedures according to the assessment of the clinical manifestation,neurological symptoms,plane of the common iliac vessels and the width of vascular window by three-dimensional computed tomography angiography between May 2004 and May 2014 were retrospectively reviewed.21 patients had undergone anterior LIF (ALIF)(13 males and 8 females with an average of 43.29±8.15 years old (range,29-53 years old);L4,5 in ten,L5S1 in 11;Meyerding grade Ⅰ 10 cases,grade Ⅱ 11 cases);25 patients had undergone posterior LIF (PLIF) (12 males and 13 females,with an average of 44.04±12.71 years old (range,25-64 years old);L4.5 in 12,L5S1 in 13;Meyerding grade Ⅰ 15 cases,grade Ⅱ 10 cases) and 24 patients had undergone transforaminal LIF (TLIF) (12 males and 12 females with an average of 45.00± 9.36 years old (range,34-62 years old);L4,5 in 12,L5S1 in 12;Meyerding grade Ⅰ 11 cases,grade Ⅱ 13 cases).Results All patients were followed up for 24-37 months (average,25.6±8.7 months).The visual analogue scale (VAS) in all groups were decreased from 7.05±0.87 (ALIF),6.60±1.39 (PLIF),6.75±1.11 (TLIF) pre-operation to 0.90±0.70,0.96±0.68,1.04±0.62 24 months after operation.Oswestry disability index (ODI) scores were decreased from 78.53％±6.25％,79.80％±6.55％,79.92％±8.10％ preoperation to 17.14％±3.01％,21.32％±4.40％,22.46％±3.87％.Perioperative data including operation time,blood loss,postoperative blood drainage,length of stay showed no significant difference between ALIF group and TLIF group,but showing significant difference comparing with PLIF group.The imageological results showed that ALIF procedure increased whole lumbar lordosis (WL) and disc height (DH),showing no significant difference comparing with PLIF group,but showing significant difference comparing with TLIF group.The ALIF procedure increased segmental lordosis angle (SL),showing significant difference comparing with PLIF group and TLIF group.The PLIF procedure decreased spondylolisthesis index (SI),showing significant difference comparing with A/TLIF group,but showing no significant difference between ALIF group and TLIF group.There was no significant difference showed comparing the L1 axis S1 distance (LASD) and sacral slope (SS),in three groups.The graft fusion was achieved in all patients after 6-8 months postoperatively without instrument failure or displacement.Conclusion Three surgical procedures of lumbar interbody fusion for adult low-grade isthmic spondylolisthesis showed silimar clinical efficacy.The ALIF procedure is superior in its capacity to restore WL,DH,and SL,and the P/TLIF procedure are superior in correcting the vertebral slippage and spinal cord decompression.
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Objective To study the effect of intra-articular injection of dehydroepiandrosterone (DHEA) on the balance of ADAMTS/TIMP-3 system in a rabbit osteoarthritis models.Methods Sixty rabbits were underwent bilateral anterior cruciate ligament transection (ACLT).Rabbits were randomizedn to the following treatment:one knee of each rabbit was treated with 100 μmol/L DHEA dissolved in dimethylsulphoxide (the experimental group) and the other knee was treated under the same schedule using dimethylsulphoxide (the control group) 4 weeks after transection,once a week for eight weeks.Twelve weeks after ACLT,all rabbits were killed after X-ray assessment and the knee joints were evaluated by gross morphology and histology.The concentration of hydroxyproline and glycosaminoglycan in the cartilage were analyzed.The mRNA expression of ADAMTS-4,ADAMTS-5,tissue inhibitor of metalloproteinases-3 (TIMP-3),transforming growth factor (TGF)-β1.Aggrecan and Collagen Ⅱ in the cartilage were analyzed using reverse transcription polymerase chain reaction (RT-PCR).The protein expression of aggrecan ARGxx and Collagen Ⅱ in the cartilage were analyzed by Western blot.Results By Mann-Whitney test,Gross morphologic scores on femoral condyle and tibial plateau in the control group were significantly higher than the experi-mental group (Z=-3.517,P＜0.01 ; Z=-2.518,P＜0.05).By unpaired Student's t test,histological evaluation showed that the grade of cartilage damage in the experimental group [(5.3±1.2) μg/ml] were less severe than that in the control group (10.1 ± 1.3,P＜0.01).The concentration of hydroxyproline [(5.7±0.3,23.6± 1.7) μg/ml] and glycosaminoglycan (30±4) in the experimental group increased significantly when compared with the control group [(4.6±0.5),(18.5±1.4),(24±4) μg/ml,P＜0.01].The mRNA expression of ADAMTS-4 (0.15±0.03)and ADAMTS-5 (0.10±0.04) in the experimental group decreased significantly compared with the control group (0.29±0.08,0.15±0.05; all P＜0.05).The mRNA expression of TIMP-3 (0.85±0.10),TGF-β1(1.2±0.4),Aggrecan (0.87±0.31) and Collagen Ⅱ (2.74±0.59) in the experimental group increased significantly when compared with the control group (0.70±0.13,0.8±0.4,0.49±0.16,2.2±0.5; all P＜0.05).The protein expression of Aggrecan ARGxx (0.53±0.10) in the experimental group decreased significantly when compared with the control group (0.81±0.12,P＜0.01).The protein expression of Collagen Ⅱ (2.3±0.7) in the experimental group increased significantly when compared with the control group (1.7±0.5,P＜0.05).Conclusion DHEA protects against cartilage degradation and inhibits the progression of OA,TGF-β1,Aggrecan and Collagen Ⅱ in cartilage may be the mechanism of the protective effect of DHEA on OA.
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Objective To observe the morphologic protection effect of intra-articular injection of osteoprotegerin (OPG)on articular cartilage in a rabbit model of osteoarthritis (OA).Methods Sixty male New Zealand rabbits were randomly divided into 3 groups:OPG group (n=20),sham-operated group (n=20) and PBS group (n=20).In OPG group and PBS group,each rabbit underwent anterior cruciate ligament transection (ACLT) in left knee joint,then 0.1 ml OPG solution or PBS were injected into the left knee for 8 weeks (5 times a week) in OPG group and PBS group,respectively.In sham-operated group,the anterior cruciate ligament was just exposed without transection,and then the incision was closed.All rabbits were sacrificed 12 weeks after operation,and the left knee joints were obtained.The Pelletier score and Mankin score were used to evaluate the macroscopic and microscopic cartilage morphology.Results The score of femoral condyle cartilage and tibial plateau cartilage was 1.80±0.89 and 1.80±0.77 in OPG group,respectively,and 3.10±0.97 and 3.20±0.77 in PBS group.However,there was no statistical difference in Pelletier score between the sham-operated group and the OPG group.Consistent with the macroscopic data,the OPG group has a significantly lower Mankin score involving cartilage structure (2.65±0.88),chondrocyte (1.35±0.71),Safranin O staining (1.83±0.83),tidemark integrity (0.30±0.47) and total score (6.13±1.97) compared with the PBS group (4.52±1.09,1.85±0.63,2.80±0.75,0.65±0.49,and 9.83±1.98,respectively).The OPG group has a significantly higher total Mankin score compared with the sham-operated group (4.80±1.25),however,there were no statistical differences in four subitem scores between the two groups.Moreover,the cartilage thickness in OPG group was 371.84±38.94 μm,255.09±74.82 μm in PBS group,and 404.68±15.97 μm in the sham-operated group,and the differences were statistically significant between three groups.Conclusion OPG has a protective effect on articular cartilage and can slow the progression of OA by reducing the grade of synovitis,decreasing the volume of osteophytes,and suppressing the loss of cartilage thickness.
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<p><b>OBJECTIVE</b>To study the interleukin-8 (IL-8) levels in gingival crevicular fluid (GCF) of porcelain teeth coated with Ni-Cr, Co-Cr or gold alloy at different time periods, and to uncover the degree of stimulation by these alloys on gingiva at different time periods.</p><p><b>METHODS</b>45 cases of porcelain teeth coated with Ni-Cr, Co-Cr or gold alloy were selected randomly, with 15 cases in each group. Sandwich enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration and total amount of IL-8 in GCF. The assay was done before treatment, as well as at 1, 3 and 6 months post-treatment.</p><p><b>RESULTS</b>The total amounts of IL-8 and GCF volume in the Ni-Cr alloy coated porcelain teeth were higher in different time period than those before treatment (P<0.05). However, the IL-8 levels in Co-Cr and gold alloy coated porcelain teeth returned to pre-dental restoration after 3 months. Otherwise, the levels of IL-8 concentration in GCF showed no significant difference among the three different alloys coating at different time periods.</p><p><b>CONCLUSION</b>IL-8 is an important cytokine during tissue inflammation. Therefore, the level of IL-8 in GCF is a useful criteria for the evaluation of stimulation degree on gingiva by different alloy coating materials in porcelain teeth. The clinical choice of different alloy coating for porcelain teeth should be considered accordingly.</p>
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Humans , Male , Alloys , Dental Porcelain , Gingiva , Gingival Crevicular Fluid , Interleukin-8 , Metal Ceramic Alloys , Tooth, ArtificialABSTRACT
Objective To discuss the formation mechanism of teardrop fracture of the axis and observe the clinical efficacy of anterior or posterior induction operation according to the fracture type.Methods Thirteen patients with teardrop fracture of the axis treated from May 2001 to October 2010 were involved in the study.There were 10 males and 3 females,at age range of 18-55 years (mean,35.5 years).Four patients were combined with cervical spinal cord injury (one patient with grade C and two with grade D according American Spinal Injury Association,ASIA).Injury causes included traffic injury in 11 patients and fall from height in two.After admission,the anteroposterior,lateral and mouth X-ray films,spiral CT sagittal reconstruction and MRI examination of the cervical vertebra were performed in all the patients.According to the injury mechanism,there were 10 patients with extension fractures treated with C2,3 intervertebral bone fusion and internal fixation through submandibular retropharyngeal approach and three with flexion fractures treated with posterior C2,3 intervertebral bone fusion and pedicle screw fixation.The clinical efficacy was also observed. Results All patients received complete exposure and effective reduction and fixation.The average time of anterior exposure and posterior exposure was 77 minutes and 125 minutes,respectively.No surgical complications occurred.All patients were followed up for 9-34 months (mean,13 months).Reexamination at four months after operation displayed bone healing in all patients.There was no lessening or breakage of the internal fixators.The spinal function of three patients with ASIA grade D injury and that of one patient with ASIA grade C injury recovered to normal at postoperative 1.5 and 6 months respectively. Conclusion The surgical approaches for teardrop fracture of the axis developed in the light of the fracture formation mechanism are beneficial and safe.
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ObjectiveTo investigate the serum soluble urokinase-type plasminogen activator (suPAR)in patients with multiple myeloma,its relationship with the 2-year survival rate and its clinical significance in predicting the disease prognosis.MethodsEnzyme-linked immunosorbent assay was applied to determine the levels of suPAR of 40 initial diagnosed patients with MM and 30 healthy persons as healthy control group.The patients were divided into three groups:effective (13 cases),remission (19 cases) and invalid (8 cases).ResultsUsually there was suPAR expression [(233.47±83.22) pg/ml] in serum of the healthy adults.There was no statistical difference of the suPAR expression in effective group or remission group before treated [(257.60±32.47) pg/ml,(331.00±99.80) pg/ml] compated with healthy group (t =2.04,t =1.83,P ＞ 0.05),and that in the invalid group,serum levels of suPAR [(562.20±291.0) pg/ml] was statistically significant with that of the effective group and that of the healthy control group (t =3.92,t =1.93,P ＜ 0.05).Through monitoring 17 patients,the suPAR of patients before treatment [(437.65±131.43) pg/ml] was statistically significant with that of healthy control group (P ＜0.01) and effective group after treatment [(298.76±108.59) pg/ml] (P ＜0.01).Serum suPAR of more than 2-year-survival patients [(333.02±85.37) pg/nl] during initial diagnosis was not statistically difference with that of the healthy control group (t =1.81,P ＞ 0.05).Serum suPAR of less than 2-year-survival patients [(646.01±103.97) pg/ml] was statistically different with that of healthy control group (t =3.84,P ＜0.01) and that of more than 2-year-survival patient group (t =3.50,P ＜0.01).ConclusionThe suPAR value was correlated to the stability of multiple myeloma,and can be used for disease prognosis and estimation of survival time.And the high expression of suPAR was related to the adverse prognosis.
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Objective The aim of the study was to explore the difference between the expression of CDg7 in peripheral blood and bone marrow on acute myeloid leukemia (AML n=30). Methods A flow cytometric quantitative analysis of expression levels for CD87 was performed on fresh blast cells in peripheral blood and bone marrow from patients with acute myeloid leukaemia using CD87, monoclonal antibodies. Analysis the difference between the expression of CD87 using matched t -test. Results The values of CD87 expression in bone marrow of 14 M5 cases are from 9.47 %～80.32 %, and from 11.49 %～87.46 % in peripheral blood. The values of CD87, expression in bone marrow of 8 M4 cases are from 14.27 %～46.28 %,and from 14.79 %～47.19 % in peripheral blood. The values of CD87 expression in bone marrow of 6 M2 cases are from 4.67 %～34.26 %, and from 8.96 %～39.78 % in peripheral blood. The values of CDs, expression in bone marrow of 2 MI cases are from 3.56 %～7.69 %, and from 5.21 %～8.96 % in peripheral blood.The expression of CD87 in peripheral blood and bone marrow from patients with acute myeloid leukaemia had statistical difference (t =3.13, P<0.05). Conclusion The levels of CD87 expression had difference between peripheral blood and bone marrow. The level in peripheral blood was higher than bone marrow. So when we performed quantitative analysis of expression levels for CD87, peripheral blood instead of bone marrow was commended.