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AIM:To investigate the role of ClC-3 chloride channel in the promotion of radio sensitization of na-sopharyngeal carcinoma CNE-2Z cells by dihydroartemisinin(DHA).METHODS:MTT was used to detect the inhibito-ry effect of DHA on the viability of CNE-2Z cells and normal nasopharyngeal epithelial NP69-SV40T cells,the radio sensi-tization effect of DHA on CNE-2Z cells was detected by cloning assay,the expression of ClC-3 protein was detected by Western blot,the expression of ClC-3 protein was down-regulated by siRNA technology,and the chlorine current of cells was recorded by whole cell patch-clamp technology.RESULTS:(1)Compared with NP69-SV40T cells,DHA selective-ly inhibited the proliferation of CNE-2Z cells,with IC10 values of(13.020±4.831)μmol/L and(5.244±1.050)μmol/L,respectively(P<0.01).(2)The results of clonal formation experiments showed that DHA had a radio sensitizing effect on CNE-2Z cells,with a radio sensitization ratio of 1.9.(3)DHA could activate the chlorine channel of CNE-2Z cells and produce an outward chlorine current,but had no effect on the chlorine channel of NP69-SV40T cells.(4)DHA promoted the expression of ClC-3 chloric channel protein in CNE-2Z cells(P<0.01).(5)Chlorine channel blocker NPPB could in-hibit the radio sensitizing effect of DHA on CNE-2Z cells by 1.84 times,and also inhibited the chlorine current activated by DHA.(6)the down-regulation of CNE-2Z ClC-3 protein could inhibit the radio sensitization effect of DHA on CNE-2Z cells by 4.19 times,and the activation of chlorine current by DHA on CNE-2Z cells was no longer produced.CONCLU-SION:DHA has a radio sensitizing effect on nasopharyngeal carcinoma CNE-2Z cells,which is likely to be related to the activation of ClC-3 chloride channel.
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OBJECTIVE@#To investigate the effectiveness of sagittal top compression reduction technique in the treatment of thoracolumbar vertebral fractures.@*METHODS@#A retrospective analysis was conducted on the clinical data of 59 patients with thoracolumbar vertebral fractures who met the selection criteria and were admitted between November 2018 and January 2022. Among them, 34 patients were treated with sagittal top compression reduction technique (top pressure group), and 25 patients were treated with traditional reduction technique (traditional group). There was no significant difference in baseline data between the two groups ( P>0.05), including gender, age, fracture segment, cause of injury, AO classification of thoracolumbar vertebral fractures, thoracolumbar injury classification and severity (TLICS) score, American Spinal Injury Association (ASIA) grading, surgical approach, preoperative vertebral body index, height ratio of the anterior margin of injured vertebra, injured vertebra angle, segmental kyphosis angle, visual analogue scale (VAS) score, and Oswestry disability index (ODI). The operation time, intraoperative blood loss, and incidence of complications between the two groups were recorded and compared. After operation, VAS score and ODI were used to evaluate effectiveness, and X-ray and CT examinations were performed to measure imaging indicators such as vertebral body index, height ratio of the anterior margin of injured vertebra, injured vertebra angle, and segmental kyphosis angle.@*RESULTS@#There was no significant difference in operation time and intraoperative blood loss between the two groups ( P>0.05). No complication such as dural sac, nerve root, or vascular injury was found during operation, and all incisions healed by first intention. Patients in both groups were followed up 6-48 months, with an average of 20.6 months. No loosening, breakage, or failure of internal fixation occurred during follow-up. The imaging indicators, VAS score, and ODI of the two groups significantly improved at 1 week and last follow-up when compared to preoperative ones ( P<0.05). At last follow-up, the VAS score and ODI further significantly improved when compared to 1 week after operation ( P<0.05). At 1 week after operation and last follow-up, the vertebral body index, segmental kyphosis angle, injured vertebra angle, and ODI in the top pressure group were significantly better than those in the traditional group ( P<0.05). There was no significant difference in VAS score and height ratio of the anterior margin of injured vertebra between the two groups at 1 week after operation ( P>0.05), but the two indicators in the top pressure group were significantly better than those in the traditional group at last follow-up ( P<0.05).@*CONCLUSION@#The treatment of thoracolumbar vertebral fractures with sagittal top compression reduction technique can significantly improve the quality of vertebral reduction, and is superior to traditional reduction techniques in relieving pain and improving spinal function.
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Humans , Thoracic Vertebrae/injuries , Lumbar Vertebrae/injuries , Retrospective Studies , Blood Loss, Surgical , Treatment Outcome , Pedicle Screws , Spinal Fractures/surgery , Kyphosis , Fracture Fixation, Internal , Fractures, Compression/surgeryABSTRACT
During pandemic of corona virus disease 2019 (COVID-19), emergency orthopedic trauma is commonly seen. It is particularly important to ensure the emergency treatment quality of orthopedic trauma but avoid cross-infection between doctors and patients. The double-buffered diagnosis and treatment mode refers to the model of patients first undergoing medical observation in the comprehensive buffer ward and the inpatient buffer rooms of various disciplines after admission to confirm the exclusion of COVID-19 and then receiving specialist diagnosis and treatment. The authors summarize the experiences of using the double-buffered diagnosis and treatment model in the Department of Orthopedics, Renmin Hospital of Wuhan University during the prevention and control of COVID-19 pandemic so as to provide a reference for treatment of orthopedic patients.
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Objective To investigate the correlation between the insertion depth of the left-sided double-lumen tube (DLT) and some specific body landmarks in order to guide left-sided DLT intubation. Methods Ninety-five adult patients who underwent thoracic surgery were chosen, and the age (A), sex (S), height (H), weight (W), distance between the cricothyroid membrane and upper notch of the sternum angle (L), size of the left-sided DLT (F), and predicted depth of intubation (y) were recorded. After anesthesia induction, the final corrected insertion depth of the left-sided DLT (Y) were recorded using fiberoptic bronchoscopy. The Y and y were compared.Linear regression and Pearson's correlation analysis were used to analyze the data. Results There was no difference between the Y and y (P> 0.05). The Y was significantly correlated with H, W, and L (P < 0.01), and was not correlated with A (P> 0.05). Three linear regression equations for H, L, and Y were obtained. H and L were linearly dependent on Y, and the determination coefficients R2 were 0.43 (Y=7.285+0.128 H) and 0.41 (Y=19.305+0.866 L), respectively. Using both H and L as the independent variables, the determination coefficient R2 was 0.56 (Y=8.127+0.087 H+0.559 L). Conclusion The linear regression equation Y=8.127+0.559 H+0.087 L could be used as a rapid method to assess the insertion depth of the left-sided DLT. However, the ideal insertion depth of the left-sided DLT still needs to be confirmed using fiberoptic bronchoscopy.
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Objective To investigate the effect of tissue engineered cartilage on the repair of artic ular cartilage defects in rabbits with calcium alginate gel (CAG) loaded transforming growth factor beta 3 (TGF-β3) and compounded with adipose mesenchymal stem cells (ADSCs).Methods The ADSCs were separated and cultured in subcutaneous fat of New Zealand white rabbits.The full-thickness articular cartilage defect model was made at the patellar groove by exposure of the femoral ankle joint.ADSCs were implanted into calcium alginate scaffolds loaded with TGF-β3 to construct tissue-engineered cartilage and transplanted into rabbit articular cartilage defects.The animals were divided into four groups:control group (injected with sterile isotonic saline),CAG group (injected with CAG),ADSCs + CAG group (injected with CAG loaded with ADSCs),TGF-β3 + CAG group (injected with CAG loaded with TGF-β3) and TGF-β3 + ADSCs + CAG group (injected with CAG loaded with TGF-β3 and ADSCs).At the end of 12 weeks,the repair of articular cartilage defects was observed by gross observation,hematoxylin-eosin (HE) staining,immunohistochemical staining of safranin-O and type Ⅱ] collagen,and the scoring method developed by In ternational Association of Cartilage Repair (ICRS) Histological score.Results The cartilage repair effect of TGF-β3 + ADSCs + CAG group was better than that of other groups,not only the neonatal tissue and the surrounding normal tissue closely,but also the secretion of extracellular matrix and normal tissue similar.The ICRS scores of each group were (7.06 ±+ 0.18) score,(7.15 + 0.23) score,(7.45 + 0.25) score,(7.47 + 0.24) score and (15.78 ±+ 0.24) score.That of TGF-β3 + ADSCs + CAG group was better than that of other groups,the difference was significant (P < 0.05).Conclusions CAG loaded with TGF-β3 and combined with ADSCs has a good effect in repairing articular cartilage defects in rabbits,and is a structural repair.
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Ovarian pregnancy is a rare ectopic pregnancy.In recent years,the development and popularization of assisted reproductive technology increase the incidence of ovarian pregnancy.This article focused on the ultrasound features of ovarian pregnancy following in vitro fertilization and embryo transfer (IVF-ET) by comparing those of pregnant corpus luteum and tubal pregnancy,and analyzed the advantages of transabdominal ultrasonography (TAS) and transvaginal ultrasonography (TVS) in the diagnosis.
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Objective To investigate the application value of DFY-Ⅱ ultrasound imaging analysis software in evaluation of fetal lung maturity in different pregnancy.Methods Totally 315 cases of healthy single pregnancies (24-41 weeks) were collected.The sonographic views of fetal right side sagittal plane were obtained.The echo intensity of fetal lung and liver were analyzed and the ratio was canculated with DFY-Ⅱ ultrasound imaging analysis software.Results The ratio of echo intensity from fetal lung and liver had positive correlation with gestational weeks (r=0.94,P<0.05),the linear regression equation was Y=0.60+0.07X (r2 =0.883).Conclusion The ratio of echo intensity by fetal lung and liver analyzed with DFY-Ⅱ ultrasound imaging analysis software can be an effective method to evaluate fetal lung maturity.
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BACKGROUND:Curcumin is proved to possess anti-inflammatory, antioxidant and anti-apoptotic roles. OBJECTIVE: To summarize and discuss the mechanisms and effects of curcumin used for the treatment of osteoarthritis METHODS: PubMed, Embase, Elseveir databases were retrieved for relevant articles published from 1973 to 2014, with the key words of “osteoarthritis, curcumin, chondrocyte, articular cartilage” in English. After the quality of the included studies was evaluated, valid data were extracted and analyzed. RESULTS AND CONCLUSION:Curcumin prevents the occurrence and development osteoarthritis by inhibiting oxidative enzyme and scavenging free radicals. Curcumin increases production of colagen type 2 through suppressing the reduction of cartilage matrix by matrix metaloproteinases. Anti-inflammatory reaction of curcumin can be achieved by inhibiting cytosolic phospholipase A2, cyclooxygenase-2, and 5-lipoxygenase. Curcumin inhibits interleukin-1β-induced apoptosis of chondrocytes and mitochondrial sweling. Results from clinical trials suggest that curcumin or its derivatives can reduce joint pain and improve the function of knee joints in patients with osteoarthritis. High-concentration curcuminin vitro have been demonstrated toxic effects.
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Objective To investigate the protective effects of carboxymethylated chitosan (CMCS) on nitric oxide (NO) induced apoptosis in rat chondrocytes, and the probable roles and mechanisms of PI3K/Akt signaling pathway in these process.Methods Rat knee articular cartilage was used as the source of chondrocytes, the cells were identified by immunohistochemical staining against collagen type Ⅱ, odium nitroprusside (SNP, 3 mmol/L) was used to establish the apoptotic models of chondrocytes.Cells were divided into four groups: the control group, the SNP-induced group, the SNP+CMCS treated group, the SNP+CMCS+PI3K inhibitor Wortmannin treated group.Cell proliferation were assessed by cell proliferation assay kit (CCK-8), the apoptotic rate of chondrocytes was determined by FCM with Annexin V-FITC/PI double staining, the expression levels of MMP-13 and TIMP-1 mRNA were detected by real-time polymerase chain reaction (PCR) analysis, the expression of Akt and p-Akt protein levels was detected by Western blotting analysis.One-way analysis of variance (ANOVA) statistical analysis was used to calculate the data.Results Three mmol/L SNP can inhibit proliferation (0.221±0.023), and the proliferation was reduced by 70% compared with the control group (0.736±0.032, F=8.203, P=0.021);and the induced apoptosis in cultured chondrocytes could be observed.The apoptotic rate was (68.8±5.2)%.Increased MMP-3 and decreased TIMP-1 mRNA expression were observed in SNP-induced cells.After adding CMCS to SNP-induced chondrocytes, the proliferation was increased while apoptotic rate was decreased, the apoptotic rate decreased to (14.7±2.3)%.CMCS could promote the activation of p-Akt in SNP-induced chondrocytes and restore SNP-induced MMP-13 and TIMP-1 mRNA expression.Conclusion CMCS could protect apoptosis in SNP induced chon-drocvtes via activation of PI3K/Akt pathwav.
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<p><b>OBJECTIVE</b>To investigate the effects of Pyrroloquinoline quinine (PQQ) on hydrogen peroxide-induced apoptosis of Schwann cells (SCs) and its mechanism.</p><p><b>METHODS</b>SCs were isolated and cultured in vitro, and identified by S-100 immunofluorescence staining. The cultured SCs were divided into control group, hydrogen peroxide-treated group, hydrogen peroxide and PQQ treated groups. The intracellular superoxide dismutase (SOD) and malondialdehyde (MDA) content was detected; the apoptotic rate of SCs induced by hydrogen peroxide was determined by flow cytometry assay. The Hoechst33342 staining was used to detect the nuclear fragmentation and apoptotic nuclear condensation of SCs; the Rhodamine123 staining was used to detect the changes of mitochondrial membrane potential in SCs, the Western blot analysis was used to detect the expression of Bcl-2 in hydrogen peroxide induced SCs.</p><p><b>RESULTS</b>The SOD activity was significantly decreased and MDA level was increased in H2O2 induced SCs (P < 0.05), after addition of PQQ, the SOD content increased and MDA content decreased (P < 0.05). Flow cytometry results showed that the early apoptotic rate was 58.8% in H2O2 induced SCs, which has significant difference compared with the control group (P < 0.05), after addition of 10, 50, 100 nmol/L PQQ, the apoptotic rates were reduced to 33.7%, 18.7%, 3.9% respectively, showing significantly different with injured group (P < 0.05). Hoechst 33342 staining showed that H2O2 induced SCs had typical morphological characteristics, such as uptake of nuclear chromatin, nuclear shrinkage, nuclear fragmentation phenomenon. The proportion of apoptotic cells after PQQ treatment reduced. Rhodamine staining results showed that the H2O2 induced mitochondrial membrane potential reduction in SCs, which was reversed by addition of PQQ. Western blot analysis showed that the expression of Bcl-2 was decreased in H2O2 induced SCs, while it increased significantly after addition of PQQ (P < 0.05).</p><p><b>CONCLUSION</b>PQQ has a protective effect on oxidative stress-induced apoptosis of SCs.</p>
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Humans , Apoptosis , Benzimidazoles , Cell Nucleus , DNA Fragmentation , Fluorescent Dyes , Hydrogen Peroxide , Pharmacology , Malondialdehyde , Metabolism , Oxidants , Pharmacology , Oxidative Stress , Pyrroles , Pharmacology , Quinine , Pharmacology , Quinolines , Pharmacology , Schwann Cells , Cell Biology , Superoxide Dismutase , MetabolismABSTRACT
BACKGROUND:Carboxymethylated chitosan is shown to promote some kinds of cells proliferation, but its effects on proliferation of Schwann cells need further studies. OBJECTIVE:To investigate the effects of carboxymethylated chitosan on proliferation of Schwann cells and expression of nuclear factor-κB in cultured Schwann cells. METHODS:Schwann cells from Sprague-Dawley rats at logarithmic growth phase were seeded in 96-wel plates, and cultured respectively with PBS, 0, 10, 50, 100, 200, 500, 1 000 mg/L carboxymethyl chitosan for 24 hours. cellproliferation was detected using the cellcounting kit-8 assay. After trypsin digestion, Schwann cells from Sprague-Dawley rats at logarithmic growth phase were used to prepare cellsuspensions, which were seeded in 6-wel cellculture plates and cultured respectively with 50, 100 and 200 mg/L carboxymethyl chitosan and PBS for 24 hours. Then, 5-bromo-2-deoxyuridine, real-time PCR and western blot assay were performed. RESULTS AND CONCLUSION:cellcounting kit-8 and 5-bromo-2-deoxyuridine detection results showed that carboxymethyl chitosan at 50-1000 mg/L, especial y at 200-500 mg/L, could promote Schwann cellproliferation. Real-time PCR and western blot results showed 50-200 mg/L carboxymethyl chitosan could promote nuclear factorκB mRNA and protein expression in Schwann cells in a dose-dependent manner, suggesting carboxymethyl chitosan can promote Schwann cellproliferation and expression of nuclear factor-κB in Schwann cells cultured in vitro.
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Objective To investigate the surgical techniques and clinical efficacy of cannulated screws combined with transposition of cuboid periosteal flap pedicled with fascia and lateral tarsal artery in the treatment of talus neck fractures.Methods From March 2008 to June 2011,12 cases with talus neck fractures were treated with cannulated screws combined with transposition of cuboid periosteal flap pedicled with fascia and lateral tarsal artery.External fixation in functional position for 12-14 weeks,and the load time was determined by X-ray fracture healing.Functional results were assessed according to AOFAS (American Orthopaedic Foot and Ankle Society) score at last follow-up.Results Twelve cases were followed up for an average of 24 months (12 to 48 months).One case of skin flap necrosis healed by dressing.All the fractures healed,and the mean healing time was 20 weeks (16 to 24 weeks).AOFAS ankle and hindfoot mean postoperative score was 82.5 (ranging from 55 to 96) points.Among them 4 cases were excellent,good in 5 cases,fair in 3 cases.Two cases presented with mild posttraumatic subtalar arthritis and pain relief after oral anti-inflammatory analgesic.One case with avascular necrosis of talus body,and X-ray displayed sclerosis of talus body,but no collapse,and the patient was told to reduce weight and regularly followed up.Conclusion The application of cannulated screws combined with transposition of cuboid periosteal flap pedicled with fascia and lateral tarsal artery for the treatment of talus neck fractures is an effective treatment,which can improves the blood supply of the talus body and reduces the occurrence of avascular necrosis of the talus.
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BACKGROUND:In the comminuted fractures of the shafts of long bones, the fracture repair process may be halted after internal fixation by locking compression plate and a higher bone nonunion rate occurs due to severe soft tissue damage, inadequate blood supply, poor cortical apposition, or unstable fracture fixation. <br> OBJECTIVE:To examine the effect of bone morphogenetic protein in the prevention of nonunion of long bone comminuted fractures after internal fixation by locking compression plate. <br> METHODS:Total y 145 long bone comminuted fracture patients were enrol ed and randomly divided into test and control groups according to the wishes of patients. The test group included 78 patients (48 males and 30 females, 18-70 years old;57 cases of closed fractures and 21 cases of open fractures) who were treated with internal fixation by locking compression plate combined with bone morphogenetic protein;the control group included 67 patients (42 males and 25 females, 18-71 years old;49 cases of closed fractures and 18 cases of open fractures) who were treated with internal fixation by locking compression plate. The fracture healing time and rate of non-union were detected and compared in the two groups after a postoperative fol ow-up of 6-18 months. <br> RESULTS AND CONCLUSION:In the test group, the average fracture healing time was 6.17 months, without non-union. In the control group, the average fracture healing time was 7.24 months, and the rate of non-union was 7%. Compared with the control group, the fracture healing time was shorter and the rate of non-union was lower in the test group (P<0.05). The use of bone morphogenetic protein is an effective method to shorten fracture healing time and prevent non-union of long bone comminuted fractures after internal fixation by locking compression plate.
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BACKGROUND:Since cannulated screw has been applied to femoral neck fracture, it is not uncommon that the screw wear penetrates or refunds. What factors affect the stability of cannulated screw for treatment of femoral neck fractures in adults remains unclear. <br> OBJECTIVE:To explore factors related to internal fixation failure by cannulated screws in treatment of adult femoral neck fracture and improve the stability of the adult femoral neck fracture by cannulated screws. <br> METHODS:A total of 92 adult patients of femoral neck fracture were treated by cannulated screws in our department between June 2007 and June 2011. Their data were retrospectively analyzed. According to clinical information and fol ow-ups, we selected factors such as age, gender, Garden type of fracture, preoperative skeletal traction, timing of surgery, Garden index, standards of pedicle screws, pedicle screw shapes, partial weight bearing time and postoperative complications, which may affect the success rate of cannulated screws for <br> treating femoral neck fracture. The selected factors were then grouped and assigned, after unrelated factors were excluded by one-way χ2 analysis, multiariable Logistic regression analysis was performed. <br> RESULTS AND CONCLUSION:The involved 92 patients were fol owed up for 18-72 months. According to Harris assessment criteria, hip function was excellent in 28 cases, good in 25 cases, fair in 17 cases, and poor in 22 cases at the final fol ow-up, the excellent and good rate was 58%. Radiographic results showed that, the patients were divided into two groups according to the presence of the displacement, GardenⅠ (n=22) and GardenⅡ (n=29) as a group, and Garden Ⅲ (n=25) and Garden Ⅳ (n=16) as the other group, the fixation failure rate was 12%and 39%, respectively. In normol and abnormal Garden Index groups, the fixation failure rate was 16%and 59%, respectively. In nail position standards and non-attainment standards groups, the fixation failure rate was 19%and 70%, respectively. In the complication and non-complication groups, the fixation failure rate was 14%and 55%, respectively. These factor groups showed significant differences (P<0.05). Multiariable Logistic regression analysis showed that, Garden type of fracture, Garden index, standards of pedicle screws, and postoperative complications are the risk factors for internal fixation failure using cannulated screws in treatment of the adult femoral neck fracture.
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Objective To study the effects of carboxymethylated chitosan (CMCS) to nitric oxide (NO)-induced apoptosis on rat chondrocytes,and explore p38MAPK signal transduction pathway in the process and its mechanism.Methods The rat articular cartilage cells were cultured in vitro,collagen type-2 (collagen-2) immunohistochemical staining was used to identify the cartilage cells.The model of chondrocyte apoptosis was built by different concentrations of sodium nitroprusside (SNP) induction.The cells were divided into the control group,the SNP treated group SNP+CMCS treated group,and the SNP+p38 MAPK inhibitor SB203580 treated group.The apoptotic rate of chondrocytes was calculated by FCM,apoptotic nuclei was identified by Hoechst33342 stain,the mitochondrial membrane potential changes was detected by Rhodamine123 (Rho123) stain,the expression of p38 and p-p38 were detected by Western blotting analysis.Results 1-3 mmol/L SNP could induce chondrocyte apoptosis,the apoptotic rate was increased with the SNP increasing,the most obvious apoptosis was occurred in 3 mmol/L SNP treated chondrocytes,which was 69.8% (P<0.05).SNP could increase the nuclear fragmentation of chondrocytes,the cells with nuclear fragmentation was significantly higher than that in the control group.SNP could reduce mitochondrial membrane potential in chondrocytes,which decreased significantly compared with the control group.SNP could increase the p-p38 expression in chondrocytes,which was 4.3 times compared to the control group.CMCS of different concentrations could reduce the apoptotic rate of SNP-induced chondrocytes,which was 51.0%,29.9% and 15.2%,which was decreased significantly (P<0.05) when compared with 3 mmol/L SNP induced group,CMCS decreased the cells number of SNP-induced nuclear fragmentation.CMCS increased the mitochondrial membrane potential in SNP-induced chondrocytes.CMCS reduced the expression levels of p-p38 in SNP-induced chondrocytes.Conclusion CMCS has protective effect on SNP-induced apoptosis of chondrocytes.This process is completed by inhibiting the activity of p38 MAPK signal pathway.
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Objective To investigate the efficacy and safety of low-dose prednisone combined with methotrexate (MTX) and hydroxychloroquine (HCQ) in the treatment of rheumatoid arthritis (RA).Methods In this 12-week study,150 patients with active rheumatoid arthritis were randomly divided into two groups:prednisone group (70 cases who were received prednisone 5 ~ 10 mg/d + MTX 10 mg/w +HCQ 0.2 g/d) and control group (80 cases who were treated by Meloxicam 7.5 mg/d + MTX 10 mg/w +Leflunomide (LEF) 20 mg/d).The primary end-points were tender and swollen joint counts,visual analogue scales (VAS),and global physician and patients assessments of disease.The secondary end-points were morning stiffness time,C-reactive protein,erythrocyte sedimentation rate,the Health Assessment Questionnaire (HAQ),DAS28 and ACR20,ACR50.Results After 12 weeks,in terms of primary endpoints,tender and swollen joint counts,VAS and global physician assessments in the prednisone group were improved significantly [(4.5 ± 2.5),(3.2 ± 3.36),(21 ± 15),(24.2 ± 16.4),(20.2 ± 10.4) vs (6.4 ±5.84),(6.6±5.5),(46±14),(37.9±19.7),(34.1±12.4),P <0.05orP <0.01].In terms of secondary end-points,the prednisone group produced higher response rates [HAQ score (0.93 ± 0.52),CRP(10.2 ± 5.8) mg/L,ESR(30 ± 14) mm/h,morning stiffness time (32.0 ± 32.3) min,DAS 28 score (3.1±0.9) vs (1.22 ±0.81),(16.3±10.1)mg/L,(33±29)mm/h,(54.7±45.4)min,(4.9±1.9),P <0.05 orP <0.01].The incidence of adverse events was similar between two groups (43% vs 49%,P > 0.05).Conclusions Low-dose prednisone combined with MTX and HCQ produced rapid and relevant improvements in RA signs and symptoms.
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Objective To observe the effect of intra-articular injection of CM-chitosan on nuclear factor κB (NF-κB) activation and nitric synthase expression in rat osteoarthritis cartilage,and to explore the mechanism of inhibition of joint destruction.Methods Thirty-six SD rats were randomly divided into three groups:A,B,C,12 in each group.Group A was the sham group,group B,C rats had the medial collateral ligaments cut off and part of medial meniscus were removed to establish osteoarthristis model.Group C rats were injected with 3% carboxymethyl chitosan intra-articularly 0.15 mg/kg 5 weeks later,and then repeated injection every 1 week.Animals were sacrificed 11 weeks after surgery.The gross changes of cartilage and the expression of NF-κB (P65) were compared by immunohistochemistry,the protein expression of I-κB and P65 in nucleus were detected by Western-bloting.Inducible nitric oxide synthase (iNOS) mRNA and protein expres-sion were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and western blot.The general score of cartilage was analyzed by H test,the rest data was analyzed by one-way ANOVA.Results The cartilage degeneration scores pf group C (intra-articular injection of CM-chitosan group) were significantly less than those of group B.The protein expression of NF-κB of the articular cartilage in group C (106±7)was significantly lower than group B (147±8),the I-κB degradation was inhibited significantly in group C,and the expression of iNOS mRNA and iNOS protein were reduced in OA art~icular cartilage of arthritis rat chondrocytes,therefore,it had protective effect on articular cartilage.Conclusion CMC may inhibit NF-κB signaling pathway by inhibiting the degradation of I-κB in cartilage,which reduces iNOS mRNA and protein expression in rat osteoarthritis cartilage,thereby protects rat osteoarthritis cartilage cells.
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Objective To evaluate the effect of free NgR-modified bone marrow stromal cell (BMSC) transplantation on axon regeneration in rats after spinal cord injury. Methods Genes encoding free NgR protein were cloned and transduced into BMSCs at passage 3 using a lentivirus vector.Indirect immunofluorescence was used to detect the expression of free NgR protein.Meanwhile a spinal cord contusion model was established in 36 adult Sprague-Dawley rats at the T10 segment.The rats were then divided randomly into an experimental group and a control group.NgR + BMSCs were transplanted into the injured site 1 week post-trauma in the experimental group.BMSCs were also transplanted at the same time into the control group.Expression of free NgR at the injury site was detected by immunohistochemical staining at 1 week post-transplantation.The functional recovery of both groups was evaluated at 4 and 6 weeks post-transplantation.Longitudinal sections of the spinal cord were studied for axon regeneration using horseradish peroxidase staining. Results Expression of free NgR was found in the cell plasma of BMSCs by indirect immunofluorescence post-transfection.Positive immunohistochemical staining for NgR was found at the transplant site in the experimental group 1 week post-transplantation.Better axon plasticity could be observed in the experimental group.The Basso-Beattie-Bresnahan scoring of the experimental group was significantly higher than that of the controls at both observation times. Conclusions Free NgR-modified BMSCs can prompt injured axons to regenerate and thus to promote the recovery of neurological function.This might provide a new strategy to treat spinal cord injury.
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Objective To investigate the effects of osteogenic growth peptide (OGP) in combination with extracorporeal shock waves (ESWs) on osteoblast proliferation.Methods Passaged cells were divided into four groups for different treatments:a control group,an OGP + ESW group,an ESW group,and an OGP group.After the respective treatments,the cells were cultured for 24 h,48 h and 72 h and counted using methylthiazdy tetrazolium (MTT) and an inverted fluorescence microscope. Immunohistochemical examination was used for detecting protein kinase A (PKA)activity,and a reverse transcription-polymerase chair reaction (RT-PCR) was used for examining PKA mRNA expression at 24 and 48 hours.Results Cell counting revealed that cell proliferation in the OGP + ESW,ESW and OGP groups was significantly promoted compared with the control group.Cell proliferation was greatest in the OGP + ESW group.The immunohistochemical examination showed positive staining intensities in the OGP + ESW,ESW and OGP groups significantly higher than in the control group.The positive staining intensity in the OGP + ESW group was again the highest.PKA activity was also significantly higher in the OGP + ESW,ESW and OGP groups than in the control group with the level in the OGP + ESW group the highest.Conclusion OGP in combination with ESW has a synergistic effect in stimulating osteoblast proliferation and growth.
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Objective To study the effect of interleukin (IL)-1β on mitochondria of chondrocytes and reactive oxygen species.Methods Rat chondrocytes were isolated and cultured in vitro,10 ng/ml IL-1β was added for establishing model of osteoarthritis.Then,ratio of apoptosis was surveyed by Annexin V-FITC and PI flow-cytometry.After Hoechst 33342 dyeing,morphology of nucleus was observed by fluorescence microscope.After rhodamine-123 luorescent staining applied,change of mitochondria membrane potential was observed by confocal microscopy.We evaluated the ability of ATP synthesizing in mitochondria by luciferase reaction.Content of active oxygen was observed by confocal microscopy.T test was used for statistical analysis.Results IL-1β could obviously reduce the mitochondrial membrane potential of chondrocytes and its ability to synthesize ATP,and could promote the expression of reactive oxygen species in cells.The values were changed to (24±4) U,(4.1±0.8) pmol/106 cells and (89±7) U from (86±10) U,(13.3±3.0) pmol/106 cells and (17±3) U.The difference had statistical significance.Conclusion IL-1β can induce chondrocyte differentiation by destroying the mitochondrial membrane integrity of chondrocytes and promoting the expression of reactive oxygen species.It can also promote the articular cartilage degeneration.