1.Technology Study of Separation and Purification of Active Constituents from Morus alba L
Degang DONG ; Xiuying ZHANG ; Xiaoxue LIU
China Pharmacy 2015;(28):3960-3963
OBJECTIVE:To optimize the technology of separation and purification of active constituents from Morus alba L. METHODS:With the adsorption rate and desorption rate as the indexes,static and dynamic macroporous resin adsorption-desorp-tion methods were adopted to optimize the type of macroporous resin,the loading amount of fluid,the volume fraction and amount of eluent in the separation and purification technology of total flavonids. With the yield and content of polysaccharide as the index-es,water extraction and alcohol precipitation method was used to optimize the volume fraction of alcohol for polysaccharide precipi-tation in the separation and purification technology of total polysaccharides. With the absorbance of total alkaloids as the index,ion exchange method was employed to optimize the loading amount of fluid and the amount of eluent in the separation and purification technology of total alkaloids. Then the verification was conducted. RESULTS:The optimal technology of total flavonids from Morus alba L. was with D101 macroporous resin as the adsorbent,loading fluid mass concentration of 0.1 g(material medicinal)/ml,loading fluid amount 9 times as the column volume(BV),70% alcohol of 5 BV for elution. The optimal volume fraction of alcohol for polysaccharide precipitation in the technology of total polysaccharides was 80%. The optimal technology of total alka-loids was with loading fluid amount of 3 BV,0.5 mol/L ammonia water of 8 BV for elution. After separation and purification,the purities of total flavonids,total polysaccharides and total alkaloids from Morus alba L. were 35.68%,47.14% and 55.79%,and the RSDs were 1.28%,1.61% and 1.14%(n=3). CONCLUSIONS:The optimal technology is stable and suitable for the separa-tion and purification of active constituents from M. alba L.
2.Effect of ClC-3 siRNA on cell cycle of HeLa cells
Dong YE ; Degang XING ; Zhi ZENG ; Lixin CHEN ; Liwei WANG
Chinese Journal of Pathophysiology 2017;33(2):257-262
AIM:To investigate the roles of ClC-3 chloride channels in the regulation of cell cycle and the re-lationship between ClC-3 chloride channels and the cell cycle regulators , such as cyclin D1, cyclin-dependent kinase (CDK)4, CDK6, P21 and P27 in the HeLa cells.METHODS:ClC-3 genes were silenced by the siRNA technique in the HeLa cells.The transfection efficiency of ClC-3 siRNA was detected by real-time PCR.The cell cycle distribution was ana-lyzed by the flow cytometry .The protein expression of ClC-3, P21, P27, CDK4, CDK6 and cyclin D1 was determined by Western blot .RESULTS:ClC-3 was knocked down by ClC-3 siRNA in the HeLa cells .Transfection of the cells with ClC-3 siRNA arrested the cells at G0/G1 phases, decreased the expression of cyclin D1, CDK4 and CDK6, and increased the expression of P21 and P27.CONCLUSION:ClC-3 plays an important role in the cell cycle of HeLa cells through the G 1-S transition point.ClC-3 may regulate the cell cycle progression by up-regulation of cyclin D1, CDK4 and CDK6 expression and/or by down-regulation of P21 and P27 expression.
3.Finite elementanalysis of clavicle fracture with superior and anterior plate fixation
Dong LIU ; Jianming WANG ; Degang ZHANG ; Kai ZHANG ; Gangqiang DU
Chinese Journal of Tissue Engineering Research 2016;20(26):3903-3908
BACKGROUND:Many scholars researched the biomechanics of middle clavicle fracture plate fixation, but little researched the plate position. OBJECTIVE:To observe the biomechanical characteristics ofanterior and superior plates for clavicle fracture with three-dimensional finite element models. METHODS:Three-dimensional finite element models of clavicle fracture with anterior and superior plates were established. The stress after anterior and superior plate fixation was analyzed. The maximum stress and displacement of plate fixation for clavicular fracture were observed under compression, torsion and three-point bending. RESULTS AND CONCLUSION:(1) In the compressed condition, the maximum stressand maximum fracture displacement were similar between the superior and anterior plate fixation (P> 0.05). (2) Under clockwise twist condition, the maximum stress and maximum fracture displacement were smaler in the superior plate fixation group than inthe anterior plate fixation group (P< 0.05). (3) Under counterclockwise twist condition, the maximum stress and maximum fracture displacement were similar between the anterior and superior plate fixation groups (P> 0.05). (4) Under three-point bending condition, the maximum stress was similar between the superior and anterior plate fixation groups (P> 0.05). The maximum fracture displacement was bigger in the superior plate fixation group than in the anterior plate fixation group (P< 0.05). (5) These findings suggest that superior fixation of clavicle fracture reconstruction plate has more advantages than the anterior plate fixation.
4.Application of Diffusion Tensor Imaging in Clinical and Basic Studies of Spinal Cord Injury (review)
Changbin LIU ; Jianjun LI ; Degang YANG ; Mingliang YANG ; Liangjie DU ; Jun LI ; Feng GAO ; Qianru MENG ; Xuechao DONG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(9):1031-1036
Diffusion tensor imaging (DTI), which responds with the diffusion of water molecules in spinal cord white matter, may be used to detect the integrity of the spinal cord fiber bundles and the pathological changes after injury. It is sensitive in acute and chronic spinal cord injury, such as cervical spondylotic myelopathy, multiple sclerosis, brain damage secondary spinal cord injury, spinal nerve root damage, and so on. In basic studies, DTI can reveal the microstructure and pathological changes of the injured spinal, and be correlated with behavioral assessment, which involved mice, monkeys, calves, cats, swines, dogs, and so on.
5.A new femoral distractor used in close reduction and internal fixation with antegrade intramedullary nail for femoral shaft fractures
Zhigang WANG ; Zhaolin WANG ; Dong LIU ; Degang ZHANG ; Long JIA ; Kai ZHANG ; Dechun ZHANG ; Mingliang MA ; Gangqiang DU
Chinese Journal of Orthopaedic Trauma 2017;19(2):164-168
Objective To introduce a novel femoral distractor which is applied in close reduction and internal fixation (CRIF) with antegrade intramedullary nail for femoral shaft fractures.Methods From September 2010 to March 2015,85 patients with femoral shaft fracture were treated by CRIF with antegrade intramedullary nail in which our self-designed novel distractor was used.They were 64 males and 21 females,with an average age of 36.6 years.By AO classification,we had 32 cases of type 32-A,40 cases of type 32-B,and 13 cases of type 32-C.The intervals between injury and surgery averaged 7.5 days (range,from 1 to 16 days).The fracture was located at the upper shaft in 26 cases,at the middle shaft in 57 cases,at the middle shaft and ipsilateral neck in one,and at the lower shaft and ipsilateral intertrochanteric site in one.Their operation time,intraoperative blood loss,intra-and post-operative complications,and fracture union time were recorded and analyzed.Results Successful close reduction was achieved in all the 85 patients with no iatrogenic injury to major vessels or nerves.Operation time averaged 105.5 minutes;time for X-ray exposure averaged 25.8 seconds;intraoperative blood loss averaged 209.2 mL.The 85 patients received follow-ups from 8 to 24 months (mean,16.3 months).Fracture union was achieved in 83 cases after an average duration of 7.8 weeks (range,from 4 to 12 weeks),but nonunion occurred in 2 cases.One-year follow-ups revealed recovery of normal function of the affected knee in all,with no limb shortening > 10 mm,rotational angulation > 15°,or lateral or anteroposterior angulation > 10°.No incision infection,deep vein thrombosis or pulmonary embolism happened during the entire follow-up.Conclusion Our self-designed novel distractor can facilitate CRIF with antegrade intramedullary nail for femoral shaft fractures,and avoid the complications and inconvenience associated with a traction table.
6.Effect of Salmon Calcitonin on Osteoporosis Induced by Spinal Cord Injury
Jun LI ; Liangjie DU ; Hongwei LIU ; Tianjian ZHOU ; Xuechao DONG ; Weijiao FAN ; Yun GUO ; Chong WANG ; Feng GAO ; Degang YANG ; Liang CHEN ; Mingliang YANG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(8):953-956
Objective To explore the effect of salmon calcitonin on osteoporosis induced by spinal cord injury. Methods 100 patients with osteoporosis induced by spinal cord injury from September 2011 to September 2014 in our department were included. They were randomly divided into control group (n=50) and observation group (n=50). The control group received vitamin D3 only, while the observation group received vitamin D3 combined with salmon calcitonin on the basis of rehabilitation physiotherapy, for 6 months. Visual Analogue Scale (VAS) of pain was evaluated in different periods. The bone mineral density (BMD) of lumbar spine and femoral neck, the parathyroid hormone (PTH), bone gla protein (BGP) and 1,25- dihydroxy vitamin D3 (1,25-(OH)2D3) were tested and recorded. Results The VAS score was lower in the observation group than in the control group 1, 2, 3 and 6 months after treatment (P<0.001). The BMD of lumbar spine and femoral neck was significantly higher, the PTH and BGP were significantly lower and the 1,25-(OH)2D3 was significantly higher in the observation group than in the control group after treatment (P<0.001). Conclusion Combination of salmon calcitonin can effectively reduce the bone pain and improve the BMD in patients with osteoporosis induced by spinal cord injury.
7.Effects of Lateral Gastrocnemius Muscle Branch Nerve Transferring on Deep Peroneal Nerve Impairment in Rats
Jun LI ; Changbin LIU ; Xuechao DONG ; Yun GUO ; Li TANG ; Liangjie DU ; Feng GAO ; Hongwei LIU ; Degang YANG ; Chong WANG ; Jianjun LI
Chinese Journal of Rehabilitation Theory and Practice 2016;22(7):779-783
Objective To explore the feasibility and effectiveness of lateral gastrocnemius muscle branch nerve transferring for deep pe-roneal nerve injury. Methods Thirty-two adult female Sprague-Dawley rats were divided into control group (n=8), sham group (n=8), nerve direct repairing group (n=8) and nerve transferring group (n=8). Twelve weeks after the anastomosis, the nerve anastomosis was observed vi-sually, the length of lateral of gastrocnemius muscle branch (L1), the diameter at the point of entering muscle (D1), the maximum detachable length of nervus peroneus communis (L2), the diameter of deep peroneal nerve (D2) and the distance between branch point and neck of fibu-la (S) were measured. The peroneal nerve functional index (PFI), the amplitude of compound muscle action potential (CMAP), nerve con-duction velocity (NCV), the weight of the tibialis anterior and the creatine kinase (CK) activity of theanterior tibial were compared among groups. Results L10.05). Conclusion It is feasible that lateral head muscular branches of gastrocnemius nerve transferring can repair deep peroneal nerve injury, which is needed to separate superficial peroneal nerve and deep peroneal nerve in the epineurium without damaging nerve for tension free neuroanastomosis. Lateral head muscular branches of gastrocnemius nerve transferring can repair the func-tion after deep peroneal nerve injury.
8.Expression of IKBKE and NF-κB in pancreatic cancer and the effect of IKBKE on proliferation and migration of pancreatic cancer cells
Jun LI ; Dong YU ; Degang KONG ; Xin LOU ; Shuang FENG ; Zhijia JIANG ; Yaqing WEI ; Ming CHEN ; Geng LIU ; Jinjin SUN
Chinese Journal of Hepatobiliary Surgery 2020;26(4):274-280
Objective:To investigate the expression of IKBKE and NF-κB in pancreatic cancer, and to explore the effect of IKBKE on pancreatic cancer proliferation and migration.Methods:Immunohistochemistry staining was used to study the expression of IKBKE and NF-κB in tissues of 61 pancreatic cancer patients admitted to the Second Hospital of Tianjin Medical University from January 2012 to January 2017 and 13 normal pancreatic tissues. The correlations between those expression to clinicopathological features were analyzed. Lentivirus mediated RNAi was transfected into pancreatic cancer cells to block IKBKE. Western blot was performed to test the silencing effeciency; CCK-8 and plate clone and scratch assays were used to investigate the proliferation and migration of pancreatic cancer.Results:Immunohistochemical staining showed that 60 (98.4%) of IKBKE staining were weakly positive, positive, and strongly positive in pancreatic cancer tissues, which were significantly higher than normal pancreatic tissues(76.9% cases were weakly positive and the rest were negative), and the differences were statistically significant ( P<0.05). All cases of NF-κB exhibited weakly positive expression and above in pancreatic cancer tissues, which was markedly higher than normal tissues (30.8% cases were weak positive and the rest were negative staining), statistically significant ( P<0.05). Survival analysis showed that patients with high level of IKBKE showed a shorter overall survival ( P<0.05). CCK-8, plate cloning and scratch assays showed that the proliferation and migration of were significantly decreased in IKBKE knocking down group ( P<0.05). Conclusions:IKBKE and NF-κB are highly expressed in pancreatic cancer, and IKBKE is correlated with NF-κB in pancreatic cancer. Blocking of IKBKE could distinctly inhibit the proliferation and migration of pancreatic cancer.