Objective To investigate the expression of transforminggrowth factor β1(TGF-β1) and insulin-like growth factorⅠ(IGF-Ⅰ)in new bone after low frequency micromovement. Methods Fifteen female sheep from Shandong province were involved in the study and their bilateral tibias transversely osteotomized in the middle shafts with a defect of 2 mm.The hind limbs were fixed with unilateral external fixators connected to a controlled micromovement device. Ten days after osteotomy, one hind limb of each sheep randomlywas selected to perform micromovement at an amplitude of 0.25 mm and a frequency of 1 Hertz, 30 min a day for 4 weeks ( micromovement group). The other hindlimb served as the control group. Five sheep were sacrificed at 3,4 and 6 weeks after osteotomy, respectively, and specimens were harvested for detecting the expression of TGF-β1 and IGF-Ⅰby immunohistochemistry and RT-PCR. Results Immunohistochemistry: In the third postoperative week in the micromovement group, the expression of TGF-β1 was detected in different areas of new chondrocytes at the margin of callus, mainly in proliferating area, and IGF-Ⅰexpressed in osteoblasts at the margin of endochondral ossification area, calcified and mature chondrocytes and osteocytes. There was seldom expression ofIGF-Ⅰ and little expression of TGF-β1 in the corresponding area in the control one. In the 4th postoperative week in the micromovement group, theexpression of TGF-β1 diminished gradually with the mature of new bone and be located in extracellular matrix and osteoblasts around ossified areas; The expression ofIGF-Ⅰ reached the peak and be located mainly in osteoblasts of new bone surface, maturing osteocytes and calcifing osteoid. But there was little expression of them in the control group. In the sixth postoperative week in the micromovement group, there was a little expression of IGF-Ⅰ expression but little expression of TGF-β1; there was nearly no expression of them in the control group. In the micromovement group, the absorbance values of TGF-β1 at 3 and 4 weeksand of IGF-Ⅰat 3, 4 and 6 weeks were significantlyhigher than those in control group（P＜0.05）. RTPCR: In the third and fourth postoperative weeks in the micromovement group, there was higher expression of mRNA of TGF-β1 and TGF-I than those in control group; in the sixth postoperative week, the expression diminished gradually, but was higher than that in control group. The absorbance values of TGF-β1 at 3 and 4 weeks and IGF-Ⅰat 3, 4 and 6weeks were significantly higher than those of control group（P＜0.05）. Conclusion Low frequency and controlled micromovement in the early stage of the fracture healing can promote the expression of TGF-β1 and IGF-Ⅰ.They worked together to regulate the process of the endochondral ossification, while in the late stage the differentiation of osteocytes and mineralization of osteoid were regulated mainly by IGF-Ⅰ, which played an important role in regulating the cell biological behavior during micromovement.
Objective To observe the effect of epidermal growth factor (EGF) on integrin alpha;5 expression and its influence on human retinal pigment epithelium (RPE) cells.Methods Human RPE cells were treated in vitro with 0.1,1.0,10.0,20.0 and 100.0 ng/ml of EGF, the mRNA and protein of integrin alpha;5 was measured by reverse transcription polymerase chain reaction（RT-PCR）and flow cytometry. Human RPE cells were cultured under 4 conditions including DMEM/F12,DMEM/F12+10 ng/ml EGF, DMEM/F12+10 ng/ml EGF+rabbit antihuman integrin alpha;5 antibody (1∶100)，DMEM/F12+10 ng/ml EGF+rabbit antihuman vimentin antibody (1∶100), and their proliferation and migration were measured by methylthiazole tetrazolium（MTT）and Boyden chamber.Results The integrin alpha;5 mRNA level of human RPE cells was not changed after 12 hours of EGF stimulation (F=0.618, P=0.687), however it was induced in a dosedependent manner after 24 and 48 hours of EGF stimulation (F=465.303, 212.340; P=0.000,0.000).The protein level of integrinalpha;5 was higher in 10 ng/ml EGF stimulation compared with the control group and 0.1 ng/ml group（P＜0.01）.MTT and Boyden chamber showed that the integrin alpha;5 expression increased the proliferation and migration of human RPE cells. Conclusion EGF can induce integrin alpha;5 expression,thus increase the proliferation and migration of human RPE cells.
Objective To observe the effect of Twist gene interference on the migration and pAkt protein expression of Rhesus retinal vascular endothelial cell line. Methods The Rhesus retinal vascular endothelial cells (RF/6A) were divided into Twist interference plasmid group, negative control group, and phosphate buffered solution (PBS) group; plasmid vectors were transfected via liposome gene transfection method. Migrated endothelial cells was detected and counted by Transwell chamber assay. Matrigel was used in endothelialcell tube formation; the inhibitory effect of Twist gene interference on endothelial cell tube formation was observed.The effect of Twist gene interference on the expression of pAkt protein in RF/6Acells was measured by Western blot. Results The number of migrated endothelial cells in Twist interference plasmid group was lower than that in the negative control and PBS group (F=23.786，P=0.000).The number of endothelial cell tubes in Twist interference plasmid group was apparently less than that in the negative control and PBS gorup (F=7.159，P=0.014). The expression of pAkt protein in Twist interference plasmid group decreased markedly.Conclusion Twist gene interference can suppress the migration of retinal endothelial cells via inhibiting the expression of pAkt protein.
ObjectiveTo investigate the effect of human subcutaneous adipose-derived stem cells (hADSCs) local transplantation on orthodontically induced root resorption (OIRR) and provide theoretical and experimental basis for the clinical application of hADSCs to inhibit OIRR. MethodsForty 8-week-old male Sprague Dawley rats were randomly divided into experimental group and control group, with 20 rats in each group, to establish the first molar mesial orthodontic tooth movement (OTM) model of rat right maxillary. The rats in the experimental group were injected with 25 μL of cell suspension containing 2.5×105 hADSCs on the 1st, 4th, 8th, and 12th day of modeling, while the rats in the control group were injected with 25 μL of PBS. The rat maxillary models were obtained before and after 7 and 14 days of force application, and 10 rats in each group were killed and sampled after 7 and 14 days of force application. The OTM distance was measured by stereomicroscope, the root morphology of the pressure side was observed by scanning electron microscope and the root resorption area ratio was measured. The root resorption and periodontal tissue remodeling of the pressure side were observed by HE staining and the root resorption index was calculated. The number of cementoclast and osteoclast in the periodontal tissue on the pressure side was counted by tartrate resistant acid phosphatase staining. Results The TOM distance of both groups increased with the extension of the force application time, and there was no significant difference (P<0.05). There was no significant difference in OTM distance between the experimental group and the control group after 7 and 14 days of force application (P>0.05). Scanning electron microscope observation showed that small and shallow scattered resorption lacunae were observed on the root surface of the experimental group and the control group after 7 days of force application, and there was no significant difference in the root resorption area ratio between the two groups (P>0.05); after 14 days of application, the root resorption lacunae deepened and became larger in both groups, and the root resorption area ratio in the experimental group was significantly lower than that in the control group (P<0.05). The range and depth of root absorption in the experimental group were smaller and shallower than those in the control group, and the root absorption index in the experimental group was significantly lower than that in the control group after 14 days of force application (P<0.05). The number of cementoclast in the experimental group was significantly lower than that in the control group after 7 and 14 days of force application (P<0.05); the number of osteoclasts in the experimental group was significantly lower than that in the control group after 14 days of force application (P<0.05). Conclusion Local transplantation of hADSCs may reduce the area and depth of root resorption by reducing the number of cementoclasts and osteoclasts during OTM in rats, thereby inhibiting orthodontic-derived root resorption.
ObjectiveTo investigate the expression of miRNA-1 in denervated skeletal muscle at different periods, and to explore effects of passive movement on the expression of miRNA-1 and differentiation of myoblasts in denervation-induced skeletal muscle atrophy in rats. MethodsTwenty-seven Sprague Dawley rats, weighing (200±10) g, were randomly divided into sham-operated group (group A, n=3), denervated group (group B, n=12), and passive movement group (group C, n=12). After the right sciatic nerve was exposed and dissociated, the sciatic nerve of 1 cm in length was removed in groups B and C; resection was not performed in group A. At 1 day after operation, passive flexion and extension movement was performed on the right hind limb in group C. At 6 hours in group A and at 3, 7, 14, and 28 days in groups B and C, 3 rats were sacrificed to measure the wet weight ratio of gastrocnemius muscle, to observe the diameter of the gastrocnemius muscle cell and evaluate the muscle atrophy by HE staining; RT-PCR was used to detect the mRNA expression of miRNA-1 and myocyte differentiation factor (MyoD), and immunohistochemistry to determine the protein expression of MyoD. ResultsAtrophy in various degrees was observed in denervated gastrocnemius muscle of groups B and C. The muscle fiber arranged in disorder and the diameter of the muscle cells decreased gradually with the time, without normal structure and morphology. The wet weight ratio and the cell diameter of the gastrocnemius in groups B and C were significantly less than those in group A (P<0.05); the wet weight ratio at 7, 14, 28 days and the cell diameter at 7, 14 days of group B were significantly greater than those of group A (P<0.05). The expressions of miRNA-1 and MyoD mRNA gradually increased with time in groups B and C, but were significantly less than those of group A at each time point (P<0.05). At 7, 14, and 28 days after operation, the expressions of miRNA-1 and MyoD mRNA in group C were significantly higher than those in group B (P<0.05). Immunohistochemical staining showed positive expression of MyoD in groups A, B, and C at each time point, but higher expression was observed in groups B and C than group A; the expression increased with time in groups B and C, and it was significantly higher in group C than group B. The correlation analysis results showed that the overall change trend of miRNA-1 and MyoD had no relation with the gastrocnemius wet weight ratio at 3 and 7 days (P>0.05), and had positive correlation at 14 and 28 days (P<0.05); positive correlation was found between the relative expression of MyoD and miRNA-1 mRNA (P<0.05). ConclusionPassive movement can prevent amyotrophy by increasing the expression of miRNA-1 and promoting the differentiation of myoblasts.
ObjectiveTo research the relevancy between the amplitudes of EMG signal of recurrent laryngeal nerve (RLN) during thyroidectemy with the movement of vocal cords after operation by applying the intraoperative neuromonitoring (IONM) and verify the proper warning criterion. MethodsFrom April 2013 to October 2013, 130 patients (214 nerves at risk) underwent complex thyroidectomy with the application of IONM. According to the degree of amplitude changing on different sites of RLN (proximal site and distal site) before closing incision, all the patients were divided into 10 groups. Every patient's vocal cords movement after operation by laryngoscopy and simulated the neural function in real time were compared. ResultsSeven patients got abnormal movement of vocal cords, the corresponding amplitudes of the EMG signal of RLN were in the range between 0 to 50%, 1 case from Group 6 (40%≤Rp/Rd<50%), 1 case from Group 8 (20%≤Rp/Rd<30%), 1 case from Group 9 (10%≤Rp/Rd<20%), 4 cases from Group 10 (0≤Rp/Rd<10%), and there's no permanent RLN palsy. ConclusionThe final amplitude of RLN decrease below 50%R1 would probably lead to vocal cords' abnormal movement, and when it decrease below 30%R1, the possibility of abnormal movement would increase; 50% decrease of EMG amplitude can be used as a warning criterion to prevent nerve function damage.
ObjectiveTo observe the effect of ultra early joint movement onthe rehabilitation of shoulder joint function in patients with breast cancer who underwent axillary lymph node dissection (ALND).MethodsA total of 100 patients with breast cancer who underwent ALND between August 2018 and December 2019 in Zhongnan Hospital of Wuhan University were randomly divided into the early movement group (n=50) and the ultra early movement group (n=50). Both groups received early rehabilitation intervention as recommended by the guidelines. Patients in the early movement group started the shoulder joint movement training on the 7th day after surgery, and patients in the ultra early movement group started the shoulder joint movement training on the 3rd day after surgery, 3 times a day, 5 days a week, for 4 weeks. The changes in pain and drainage volume 3 days, 1 week, and 2 weeks after surgery and the changes of shoulder joint range of motion 1 week, 2 weeks , and 3 weeks after surgery were compared between the two groups, changes in shoulder function and quality of life 1 week and 3, 6, and 12 weeks after surgery were compared by the Constant-Murley and the Medical Outcomes Study 36-item Short-form Health Survey (SF-36) scales, respectively.ResultsThree days, 1 week, and 2 weeks after surgery, no significant difference in the pain scores or drainage volumes was observed between the two groups (P>0.05). One week, 2 weeks, and 3 weeks after operation, the motion ranges of shoulder abduction, flexion, and external rotation in the ultra early movement group were significantly better than those in the early movement group (P<0.05), and the motion range of shoulder internal rotation 1 week after operation in the ultra early movement group was significantly better than that in the early movement group (P<0.05). One week and 3, 6, and 12 weeks after operation, the Constant-Murley scores in the ultra early movement group were 25.9±4.3, 55.4±5.3, 64.6±4.5, and 73.3±4.6, respectively, which were better than those in the early movement group (21.3±3.8, 48.9±7.8, 57.3±4.7, and 70.7±3.0, respectively; P<0.05). No significant difference in the SF-36 scale scores was observed between the two groups (P>0.05).ConclusionsUltra early joint movement can significantly improve the motion range and functions of shoulder joint in patients with breast cancer who underwent ALND. What’s more, ultra early joint movement does not increase the early drainage volume or pain, and has no significant impact on the later quality of life. It is worthy of clinical application.
ObjectiveTo review the current status and progress of locking plate for the treatment of distal femoral comminuted fractures.MethodsThe related literature was extensively reviewed to summarize the current status and progress in the treatment of distal femoral comminuted fracture with locking plate from four aspects: the current treatment situation, the shortcomings of locking plate and countermeasures, the progress of locking technology, locking plate and digital orthopedic technology.ResultsTreatment of distal femoral comminuted fractures is challenging. Locking plates, the most commonly used fixation for distal femoral comminuted fractures, still face a high rate of treatment failure. Double plates can improve the mechanical stability of comminuted fractures, but specific quantitative criteria are still lacking for when to choose double plates for fixation. The far cortial locking screw has shown good application value in improving the micro-movement and promoting the growth of callus. The biphasic plating is a development of the traditional locking plate, but needs further clinical examination. As an auxiliary means, digital orthopedic technology shows a good application prospect.ConclusionThe inherent defect of locking plate is a factor that affects the prognosis of distal femoral comminuted fracture. The optimization of locking technology combined with digital orthopedic technology is expected to reduce the failure rate of treatment of distal femoral comminuted fracture.
Objective To explore the effect of bone morphogenetic protein 4 (BMP4) on the glycolysis level of human retinal microvascular endothelial cells (hRMECs). MethodsA experimental study. hRMECs cultured in vitro were divided into normal group, 4-hydroxynonenal (HNE) group (4-HNE group) and 4-HNE+BMP4 treatment group (BMP4 group). 4-HNE group cell culture medium was added with 10 μmmol/L 4-HNE; BMP4 group cell culture medium was added with recombinant human BMP4 100 ng/ml after 6 h stimulation with 10 μmol/L 4-HNE. The levels of intracellular reactive oxygen species (ROS) were detected by flow cytometry. The effect of 4-HNE on the viability of cells was detected by thiazole blue colorimetry. Cell scratch test and Transwell cell method were used to determine the effect of 4-HNE on cell migration. The relative expression of BMP4 and SMAD9 mRNA and protein in normal group and 4-HNE group were detected by real-time quantitative polymerase chain reaction and Western blot. Seahorse XFe96 cell energy metabolism analyzer was used to determine the level of intracellular glycolysis metabolism in normal group, 4-HNE group and BMP4 group. One-way analysis of variance was used for comparison between groups. ResultsThe ROS levels in hRMECs of normal group, 4-HNE group and BMP4 group were 21±1, 815±5, 810±7, respectively. Compared with the normal group, the levels of ROS in the 4-HNE group and the BMP4 group were significantly increased, and the difference was statistically significant (F=53.40, 50.30; P＜0.001). The cell viability in the normal group and 4-HNE group was 1.05±0.05 and 1.28±0.05, respectively; the migration rates were (0.148±0.005)%, (0.376±0.015)%; the number of cells passing through the pores were 109.0±9.6, 318.0±6.4, respectively. Compared with the normal group, the 4-HNE group had significantly higher cell viability, cell migration rate, and the number of cells passing through the pores, and the differences were statistically significant (F=54.35, 52.84, 84.35; P＜0.05). The relative expression levels of BMP4 and SMAD9 mRNA in the cells of the 4-HEN group were 1.680±0.039 and 1.760±0.011, respectively; compared with the normal group, the difference was statistically significant (F=53.66, 83.54; P＜0.05). The relative expression levels of BMP4 and SMAD9 proteins in the cells of the normal group and 4-HEN group were 0.620±0.045, 0.860±0.190, 0.166±0.049, 0.309±0.038, respectively; compared with the normal group, the differences were statistically significant (F=24.87, 53.84; P＜0.05). The levels of intracellular glycolysis, glycolytic capacity and glycolytic reserve in normal group, 4-HNE group and BMP4 group were 1.21±0.12, 2.84±0.24, 1.78±0.36, 2.59±0.11, 5.34±0.32, 2.78±0.45 and 2.64±0.13, 5.20±0.28, 2.66±0.33. Compared with the normal group, the differences were statistically significant (4-HNE group: F=86.34, 69.75, 58.45; P＜0.001; BMP4 group: F=56.87, 59.35, 58.35; P＜0.05). There was no significant difference in intracellular glycolysis, glycolysis capacity and glycolysis reserve level between 4-HNE group and BMP4 group (F=48.32, 56.33, 55.01; P＞0.05). ConclusionBMP4 induces the proliferation and migration of hRMECs through glycolysis.
When performing eye movement pattern classification for different tasks, support vector machines are greatly affected by parameters. To address this problem, we propose an algorithm based on the improved whale algorithm to optimize support vector machines to enhance the performance of eye movement data classification. According to the characteristics of eye movement data, this study first extracts 57 features related to fixation and saccade, then uses the ReliefF algorithm for feature selection. To address the problems of low convergence accuracy and easy falling into local minima of the whale algorithm, we introduce inertia weights to balance local search and global search to accelerate the convergence speed of the algorithm and also use the differential variation strategy to increase individual diversity to jump out of local optimum. In this paper, experiments are conducted on eight test functions, and the results show that the improved whale algorithm has the best convergence accuracy and convergence speed. Finally, this paper applies the optimized support vector machine model of the improved whale algorithm to the task of classifying eye movement data in autism, and the experimental results on the public dataset show that the accuracy of the eye movement data classification of this paper is greatly improved compared with that of the traditional support vector machine method. Compared with the standard whale algorithm and other optimization algorithms, the optimized model proposed in this paper has higher recognition accuracy and provides a new idea and method for eye movement pattern recognition. In the future, eye movement data can be obtained by combining it with eye trackers to assist in medical diagnosis.