BACKGROUND:Whether continuous passive motion improves osteoarthritis by enhancing the proliferation ability of chondrocytes is rarely reported.
OBJECTIVE:To analyze the therapeutic outcomes of continuous passive motion in rabbits with osteoarthritis and the underlying mechanism.
METHODS:Thirty-six New Zealand white rabbits were randomly al otted into three groups (n=12 per group). Rabbits in control group only underwent capsulotomy with no harm to the cartilage;osteoarthritis models were established in the rabbits of model and treatment groups using Hulth method. At 1 day after modeling, the treatment group rabbits were treated with continuous passive motion, 8 hours daily for consecutive 8 weeks. Interleukin-1 and tumor necrosis factorαlevels in the synovial fluid were detected by ELISA;col agen type II expression and the proliferation ability of chondrocytes were detected by MTT assay;Erk signaling pathway activation was determined using western blot assay.
RESULTS AND CONCLUSION:In the model group, interleukin-1 and tumor necrosis factorαlevels in the synovial fluid were significantly increased, and the expression level of col agen type II mRNA was remarkablely down-regulated. Continuous passive motion significantly downregulated interleukin-1 and tumor necrosis factorαlevels and up-regulated the col agen type II mRNA level (P<0.01). The model group showed significantly decreased proliferation ability of chondrocytes and down-regulated Erk signaling pathway activation, while after continuous passive motion, al above indicators were significantly improved (P<0.01). These results indicate that the continuous passive motion can al eviate osteoarthritis probably by influencing interleukin-1 and tumor necrosis factorαlevels, proliferation ability of chondrocytes, and col agen type II expression, as wel as regulating Erk signaling pathway activation.

Objective To explore the association between the polymorphism rs671 site of the acetaldehyde dehydrogenase 2 (ALDH2) gene and H-type hypertension in the elderly in Han nationality in Qingdao.Methods Totally 406 patients aged 60-90 years with primary hypertension were randomly selected in the study.Serum levels of homocysteine(Hcy),folate and vitamin B12 were determined and all patients were divided into H-type hypertension group and non H-type hypertension group.Gene chip technology was used to analyze the ALDH2 (rs671) polymorphism,and the association between ALDH2 gene and H type hypertension was evaluated.Results Of all hypertensive participants,82.0％ (333/406) were in H-type hypertension,87.4％ (221/253) in male and 73.2％ (112/153) in female.The GA/AA genotype and A allele frequency were higher in H-type hypertension group than in non H-type hypertension group [37.2 ％ (124/333) vs.16.4 ％ (12/73) and 21.3％(71/333) vs.9.6％(7/73),P=0.001and 0.021].Serum Hcyleveland the prevalence of H type hypertension were higher in GA/AA genotype group than in GG genotype group(both P＜ 0.05).Logistic regression analysis showed that GA/AA genotype,gender (male),drinking,folate deficiency and increased systolic pressure were the risk factors for H-type hypertension (OR=3.17,2.14,2.37,0.75,1.03,respectively,all P＜ 0.05).Conclusions The genetic variation of ALDH2(rs671) may increase Hcy level by decreasing the levels of folate and vitamin B12.GA/AA genotype is a risk factor for H-type hypertension,and it contributes to H-type hypertension together with gender,drinking history,folate levels and systolic pressure.

Objective To explore the clinical efficacy of medial iliosciatic plating via the Stoppa approach for complex acetabular fractures involving the posterior column.Methods Between February 2015 and February 2016,a total of 16 complex acetabular facture cases treated by the medial iliosciatic plate via the Stoppa approach were retrospectively analyzed in this study.This approach provided good exposure to a large region of the pelvis and acetabulum including pubis symphysis,pubic ramus,anterior and inner wall of acetabulum,quadrilateral surface,inner surface of posterior column,true pelvic margin,greater sciatic notch and sacroiliac articulation.The anterior and column was reduced and fixed by the anterior column plate and the medial ilioseiatie plate.The screw direction and angle were adjusted according to the intraoperative X-ray.Surgical time,amount of bleeding,and relevant complications were recorded.The reduction of the posterior column fracture was evaluated by Matta scoring system on the plain X-ray of the pelvic post-surgery,and functional outcomes of the hip joint affected were evaluated one year post-surgery by the Merle d'Aubigne-Postel scoring system.All the cases were followed for at least 12 months.Results The reduction and fixation of the posterior column was accomplished in all the 1 6 patients.The average surgical time was 165.5 min (range,130-270 min).The average blood loss was 1 245.6 ml (range,600-5 600 ml).Thc intraoperative infusionof concentrated red blood cells averaged 6 units.According to the Matta scoring system,anatomical reduction was achieved in 12 cases,satisfactory reduction in 3,and poor reduction in one.The patients were followed from 12 to 22 months.According to the Merle d'Aubigne-Postel scoring system,there were 11 cases of excellent and 3 cases of good,yielding a good or excellent rate of 87.5％.The average Merle d'Aubigne-Postel score was 15.8 (range,8-18).There were 1 case of external iiiac vein rupture and 1 case of bladder rupture.Both were repaired during surgery.Superior gluteal artery rupture was found in 1 case and surgical ligation of the artery was performed during surgery.Conclusion In the treatment of complicated acetabular fractures involving the posterior column,the medial iliosciatic plating via the Stoppa approach is safe and effective,because it can provide a safe and sufficient operative field for surgeons to reduce and fix the posterior column fractures,and it leads to satisfactory recovery of the patients with limited complications.

Objective To prepare the human papillomavirus (HPV) 16 peptide vaccine and explore the effect in vitro and in vivo. Methods (1) Prediction of the major histocompatibility complex (MHC) class I restricted T cell epitopes by bioinformatics target at transporter associated with antigen processing (TAP) and named by E7Pa, E7Pb, E7Pc separately. (2)In vivo, the C57BL/6 mice were divided into five groups with same amounts randomly after loading with TC-1 cells (HPV 16 positive tumor cells from C57BL/6 mouse), named as E7Pa + CpG,E7Pb + CpG,E7Pc + CpG (as experiment groups, and added 50 μg/ml E7Pa, E7Pb, E7Pc, respectively), CpG(as positive control group and added Con A with 12 mg/L final concentration) and blank control group (without any treatment). The T cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay at different time points;the lactate dehydrogenase (LDH) delivery method was used to test the cytolytie T lymphocyte (CTL) activity of mouse splenic lymphocyte in different ratio of effector cells and target cells (E:T);the related cytokines in tumor tissue and mouse peripheral blood were evaluated by real-time PCR and enzyme-linked immunosorbent assay (ELISA), respectively. The tumor volumes were measured to contrast the therapeutic effect in different groups. Results (1) Three peptide named E7Pa, E7Pb, E7Pc were successfully preparated which had high affinity and specificity. (2) After vaccination of 24, 48, 72,96 hours, MTT results shown that the proliferation rate in E7Pa + CpG group were(131±32)%, (302±15)%, (552±28)%, (731±24)% individually, which were much higher than those in blank control [(72± 15) %, (120 ± 57) %, (176 ±41)%, (288±29)% ;P<0.01], and the other groups i. e. E7Pb + CpG,E7Pc +CpG and CpG groups all proliferated much higher than those in blank control group with statistic signification (P<0. 05), but there was no significant difference between groups(P>0.05);the LDH delivery assay showed that when the ratio of E:T was 100:1, the activity of CTL in the E7Pa + CpG group was most powerful than the other groups with statistic signification (P<0. 01). Meanwhile, the ratio of E:T was concentration-dependent. Compared E7Pb + CpG, E7Pc + CpG or CpG groups with blank control group, there were significantly difference(P<0. 05) ,while there was no significant difference between groups(P >0. 05). The mRNA levels of interferon γ (IFN-γ), interleukin-2 (IL-2) in tumor tissue and peripheral blood in E7Pa + CpG group were significantly higher than those in blank control group (P<0. 01), which was the similar results when compared E7Pb + CpG, E7Pc + CpG or CpG groups with control group (P < 0. 05), and without significant difference between groups(P > 0. 05). The tumor volumes were suppressed obviously in all the experiment groups, especially at the 60th days, the volumes in ETPa + CpG group were much smaller than that in blank control group with statistic signification (P < 0. 01),which was the similar results that E7Pb + CpG, E7Pc + CpG or CpG groups had difference than blank control group with statistic signification (P < 0. 05), and without significant difference between groups(P >0. 05). Conclusion The HPV16 E7 peptide target at TAP combination with CpG as a vaccine could treat effectively the HPV16 E7 positive tumor in experiment.

Objective To observe the change of MR perfusion value in patients with hepatocellular carcinoma (HCC) before and after transcatheter arterial chemoembolization (TACE). Methods A total of 22 patients with HCC underwent MR perfusion weighted imaging (MR PWI) before TACE and 3-10 days after TACE. The mean time to enhance (MTE), negative enhancement integral (NEI), time to peak (TTP) and maximum slope of decrease (MSD) before and after TACE were acquired and compared. Results The time intension curve (TIC) of HCC region was observed to descend rapidly before TACE, while descended slowly after TACE. The value of MTE and TTP after TACE were lower than those before TACE (P<0.05), and the value of NEI after TACE was higher than that before TACE (P<0.05). The value of MSD after TACE were lower than that before TACE, but no statistical significance was found (P>0.05). Conclusion MR PWI is a very sensitive imaging technique that be used to monitor blood flow changes of HCC before and after TACE and evaluate efficacy of TACE.

OBJECTIVETo study the acute effects of dimethoate on the muscarinic-receptors(M1, M2) in the brain of rats.

METHODS24 Sprague-Dawley rats were divided into 4 groups randomly. They were administered subcutaneously with 0, 25, 50, 100 mg/kg dimethoate, respectively. Brains were removed after 48 hours of administration. Radioligand binding assay was used to determine the density and affinity of M1 and M2 receptors.

RESULTSRats in the treated group showed low density of M1 and M2 receptors compared with the control rats. The brain M1 receptor density of the rats in the highest dosage group was significantly lower than that in the control group while brain M2 receptors density had a decrease trend with increasing dosage, but the difference showed no significance. However, there were no differences of the affinity of both M1 and M2 among different treated groups. Correlation analysis showed there is positive relationship between cholinesterase activity and density of M1 receptors(r = 0.583, P < 0.01).

CONCLUSIONM1 and M2 receptors density decreased with the increasing dosage of dimethoate. It is suggested that the alleviating of cholinergic symptoms may be due to the decrease of M1 and M2 receptors in rat brain.

Animals ; Brain ; drug effects ; metabolism ; Cholinesterases ; metabolism ; Dimethoate ; pharmacology ; Dose-Response Relationship, Drug ; Radioligand Assay ; Rats ; Rats, Sprague-Dawley ; Receptor, Muscarinic M1 ; analysis ; drug effects ; Receptor, Muscarinic M2 ; analysis ; drug effects

The effect of magnetic stimulation (MS) on sciatic nerve injury was observed. After sciatic nerve was crushed in 40 Sprague Dawley (SD) rats, one randomly selected group (group D) was subjected, from the 4th day post-operatively to 3 min of continuous 70% of maximum output of MS daily for 8 weeks. The other group (group E) served as a control group. The nerve regeneration and motor function recovery were evaluated by walking track analysis (sciatic function index, SFI; toe spreading reflex, TSR), electrophysiological, histological and acetylcholineesterase histochemistry. The SFI in the group D was greater than in the group E with the difference being statistically significant (P < 0.01). TSR reached its peak on the 4th day in the group D and on the 10th day in the group E respectively. The amplitude and velocity of MCAP and NCAP in the group D was greater than in the group E with the difference being statistically significant (P < 0.01), while the latency and duration of MCAP and NCAP in the group D were less than in the group E with the difference being also statistically significant (P < 0.01). Histological examination showed the mean axon count above the lesion for thick myelinated fibers (> 6.5 microns) in the group D was greater than in the control group with the difference being statistically significant (P < 0.01), while the mean axon count below the lesion for thick myelinated fibers was less than that in the group E with the difference being statistically significant (P < 0.01). The mean axon count above the lesion for thin myelinated fibers (2-6.5 microns) in the group D was greater than that in the group E with the difference being statistically significant (P < 0.05), while the mean axon count below the lesion for thin myelinated in the group D was greater than that in the group E with the difference being statistically significant (P < 0.01). Acetylcholine esterase examination showed that the MS could significantly increase the number of the motor neurons. There was no significant difference in the number of the motor neurons between the treatment side and the normal side (P > 0.05). It can be concluded that MS can enhance functional recovery and has a considerable effect in the treatment of the peripheral nerve injury.
Acetylcholinesterase/metabolism ; Electromagnetics ; Motor Neurons/physiology ; *Nerve Regeneration ; Random Allocation ; Rats, Sprague-Dawley ; Sciatic Nerve/*injuries ; Sciatic Nerve/*physiopathology ; Sciatic Neuropathy/rehabilitation

Transforming growth factor-beta (TGF-beta) may cause cell cycle arrest, terminal differentiation, or apoptosis in most normal epithelial cells, whereas most malignant cell lines are resistant to TGF-beta. Mechanisms of resistance to TGF-beta caused by modulation of cell cycle regulators and/or inactivation of components of the TGF-beta signaling transduction pathway such as C-myc and Smad4 are not well understood. To investigate the potential association between loss of sensitivity to TGF-beta and expression status of transforming growth factor receptor II (TbetaR II) , Smad4, CDC25A and C-myc in 14 cell lines derived from ovarian cancer, the expression levels of these genes were detected by semi-quantitative RT-PCR. Normal ovarian surface tissues were used as controls. The expression of TbetaR II was detectable in all of 14 cell lines. The expression of Smad4 was decreased in 10 cell lines and 9 cell lines overexpressed CDC25A, as compared to normal controls. CDC25A gene was overexpressed with 88% (8/9) in tumorigenic cell lines as determined by xenografts in nude mice, and only in 20% (1/5) of non-tumorigenic cell lines (P<0.05). C-myc was not overexpressed in any of these cell lines. The loss of sensitivity to TGF-beta of cell lines derived from ovarian cancers may be related to a decreased expression of Smad4, which mediates TGF-beta induced growth inhibition, and/or an overexpression of CDC25A. This overexpression of CDC25A correlates with increased tumorigenicity of ovarian cancer cell lines. The loss of sensitivity to TGF-beta is not associated with a lack of TbetaR II.
Animals ; Cell Line, Tumor ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Mice ; Mice, Nude ; Ovarian Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-myc ; metabolism ; Receptors, Transforming Growth Factor beta ; metabolism ; Signal Transduction ; Smad4 Protein ; Trans-Activators ; metabolism ; Transforming Growth Factor beta ; pharmacology ; cdc25 Phosphatases ; metabolism

Objective To clarify the function of LIM and SH3 domain protein-1 (LASP1) and ferritin in rhBMP2-induced osteogenic differentiation of beagle bone marrow mesenchymal stem cells (BMSCs).Methods After BMSCs from 3-18-month-old C57BL/6J mice were cultured adherently for 24 hours,they were subjected to osteogenic differentiation for 7,14 and 21 days in 3 groups.BMP2 (100 μg/L) and osteogenic differentiation medium was added in the experimental group,only osteogenic differentiation medium was added in the control group,and nothing was added in the blank group.Osteoblast differentiation was determined by examining marker genes (Runx2,OSX,OCN and OPN) using qRT-PCR.The protein expression of both LASP1 and ferritin was investigated using western blotting.After LASP1 and ferritin were silenced in the cells in the experimental group after transfection of shRNA to target LASP1(m),rhBMP2-induced osteogenesis was repeated to verify the roles of LASP1 and ferritin in osteoblast differentiation.Results The qRT-PCR showed successful osteoblast differentiation in the experimental group.Western blotting verified significant down-regulation of LASP1 and up-regulation of ferritin in the experimental group.After the LASP1 gene was silenced,the expression levels of osteoblast differentiation marker genes in the experimental group were higher than those in the control group.Conclusions rhBMP2 can induce mouse BMSCs to differentiate into osteoblasts in a significant manner.Combined with our preliminary research,the present study may confirm that LASP1 and ferritin,which play an important role in regulating cytoskeleton activity and iron metabolism,are critical in the osteogenic differentiation of mouse BMSCs induced by rhBMP2.

The influence of pulsed magnetic stimulation (MS) on the sciatic nerve injury was investigated. Thirty rats were divided into three groups equally: MS group (A), electric stimulation (ES) group (B) and the control group (C). The MS and ES were applied immediately after the first 10 min of the sciatic nerve crush. Sciatic function index (SFI), toe spreading reflex (TSR), muscular weight and volume were measured after the experiment. The TSR of in the groups A and B occurred at 4th day while in the control group it occurs at 10th day. There was statistically significant difference in SFI between groups A and B (P<0.01). The weight and volume of the gastrocnemius muscle were statistically greater in the groups A and B than in the control group (P<0.01). The effect of MS was similar to that of ES. It was suggested that the application of MS immediately after the nerve injury might have an important clinical value as it can accelerate functional recovery and prevent or minimize muscle atrophy. The technique is easily to operate, non-invasion, painless and permits tolerance of high intensity output to be used.