Corn meal was hydrolyzed in HCI ,and pH adjusted to 6.8 with (NH_4)OH. The substrate was inoculated with Cadida trapicalis ,then cultivated in a shaker at 30℃ and in 200r/min for 2 days. The Results indicated that the protein content in corn meal grew from 8.4 to 27.4%the lysine in the protein increased from 32.1 to 73.48/kg,the tryptophan from 6.8 to to 14.8g/kg. This shows that the nutritional value of corn meal can be improved with this method.

Objective To study the quality control of previous process of apheresis platelets detection .Methods Single and double apheresis platelets each of 100 samples were collected and were diluted 1∶1 ,1∶3 and 1∶7 .Aautomatic Blood Cell Count-er was employed to detect the platelet count .Another 100 samples of apheresis platelets were collected and stand at room tempera-ture for 0 ,30 ,60 ,90 ,120 min ,After 1∶3 dilution ,the platelet counts were detected .Results Differences of platelet counts be-tween 1∶1 and 1∶3 dilution ,1∶3 and 1∶7 dilution of single or double apheresis platelets showed statistically significant differ-ences(P<0 .05) .Differences of platelet counts between standing for 0 ,30 min and standing for 60 ,90 ,120 min were found statis-tically significant(P<0 .05) .Conclusion Quality control of previous process of apheresis platelets detection is very important for platelet count after collection .

10g corn meal was mixed with 200ml Na2HPO4/KH2PO4 buffer (1/15M, pH6.81, containtng ammonium oxalate 0.6%, MgSO4 0.01%, FeSO4 0.002%) in a 500ml flask, then was inoculated with Candids tropicalis, and 1.5g ?-amylase was added at the same time. The substrate was cultivated in a shaker at 30℃, 200r/min for 3 days. The protein content increased from 8.4% to 23.4% (P

Objective To investigate the mechanisms of apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase. Methods Human leukemia cell line K562 were exposed to indole-3-acetic acid (IAA) at 20, 40, 60, 80 or 100 mol/L and horseradish peroxidase(HRP) at 1.2 g/mL for varying times. MTT assay was applied to detect the cell proliferation. Flow cytometry was performed to detect the arrest of cell cycle. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to measure apoptosis. 2, 7-dichlorofluorescin diacetate (DCFH-DA) uptake was measured to determine free radical by confocal microscope. Content of malondiadehyde (MDA) and activity of superoxide dismutase (SOD) were measured by biochemical methods. Results IAA/HRP initiated growth inhibition of K562 cells in a dose- and time-dependent manner. Flow cytometry revealed that cell cycle arrested at G1/G0 after 24 hours treatment. After 72 hours treatment, apoptotic rate of 100 mol/L IAA group increased to 43.9%, which was 5 times that of control(P＜0.01). Content of MDA and activity of SOD increased respectively in treatments compared with control. Meanwhile, IAA/HRP stimulated the formation of free radical, which was increased by IAA concentration-dependently. Conclusion The combination of IAA and HRP can inhibit the growth of Human leukemia cell line K562 in vitro by inducing apoptosis which is associated with the increase of free radical. The combination of IAA and HRP might be a promising chemopreventive and chemotherapeutic agent against human leukemia.

OBJECTIVETo construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressing HKE-HERG-G572R.

METHODSHERG-G572R mutant fragment was constructed by over-lap extension PCR and validated by DNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cell line stably expressing HKE-HERG-G572R.

RESULTSThe pcDNA3-HERG-G572R expression vector was successfully constructed and the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-fold HKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfected with pcDNA3 (P<0.01).

CONCLUSIONThe protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to provide a cell model for studying individualized therapy.

Base Sequence ; Ether-A-Go-Go Potassium Channels ; genetics ; Gene Expression ; Genetic Vectors ; HEK293 Cells ; metabolism ; Humans ; Mutation ; Transfection

Objective To construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressing HKE-HERG-G572R. Methods HERG-G572R mutant fragment was constructed by over-lap extension PCR and validated by DNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cell line stably expressing HKE-HERG-G572R. Results The pcDNA3-HERG-G572R expression vector was successfully constructed and the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-fold HKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfected with pcDNA3 (P<0.01).Conclusion The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to provide a cell model for studying individualized therapy.

Objective To construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressing HKE-HERG-G572R. Methods HERG-G572R mutant fragment was constructed by over-lap extension PCR and validated by DNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cell line stably expressing HKE-HERG-G572R. Results The pcDNA3-HERG-G572R expression vector was successfully constructed and the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-fold HKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfected with pcDNA3 (P<0.01).Conclusion The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to provide a cell model for studying individualized therapy.

Objective To investigate the effect of facet joint resection at different ranges under endoscopy on the stability of the cervical spine and provide a biomechanical theoretical basis for clinical surgery.Methods A normal finite element model of the cervical spine C5-6 was established based on CT data,and unilateral facetectomy models with different ranges(0,25％,50％,75％,and 100％)of laminectomy were obtained(Models 1-5)by simulating cervical endoscopic surgery.The ranges of motion(ROMs)of C5-6 and the von Mises stresses of the discs for the models in each group were compared and analyzed.Results Except for flexion,Models 1 and 2 showed insignificant changes in ROMs and disc von Mises stresses in each direction compared with those of the normal model.Model 3 showed a noticeable increase in ROMs and disc von Mises stresses in each direction compared with those of the normal model:ROMs under flexion,extension,left lateral bending,right lateral bending,left rotation,and right rotation increased by 27％,4％,3％,13％,5％,and 16％,respectively,and von Mises stresses increased by 32％,4％,2％,5％,9％,and 5％,respectively.Models 4 and 5 exhibited a significant increase in the ROMs and disc von Mises stresses in each direction compared to the normal model.For Model 4,ROMs were increased by 27％,14％,6％,24％,7％,167％,and von Mises stresses were increased by 33％,13％,3％,32％,10％,130％.For Model 5,ROMs were increased by 27％,17％,6％,25％,7％,167％,and von Mises stresses were increased by 33％,29％,8％,33％,12％,138％.Conclusions As the range of unilateral facetectomy increased,cervical ROM and disc von Mises stress extremum gradually increased.The cervical spine shows a significant ROM increase and stress changes when facet joint resection on one side exceeds 1/2.More than 1/2 of the facet joint should be preserved during surgery to avoid medical instability.

BACKGROUND:In clinical application,simple interspinous fixation without additional interbody fusion has similar fixation effects to pedicle screw and rod fusion internal fixation,and can effectively reduce the range of motion of the responsible segment and the stress of the articular process.However,after simple placement of the new interspinous fusion fixation device BacFuse,the stress at the root of the spinous process is relatively concentrated,and the spinous fracture is prone to occur.If an intervertebral fusion cage is inserted in conjunction with interspinous fixation,Von Mises stress can theoretically be dispersed to reduce the risk of spinous fracture.However,there are few studies on biomechanics and finite element analysis. OBJECTIVE:To observe the biomechanical stability of interspinous fixation-assisted endoscopic interbody fusion in the treatment of severe lumbar spinal stenosis. METHODS:The normal finite element model M0 of the L4-L5 segment of the lumbar spine was established by Mimics,Geomagic,Solidworks,and ANSYS software based on the lumbar CT images of a 26-year-old adult male volunteer excluding spinal diseases.On the basis of M0,the immediate model M1 after endoscopic decompression combined with interbody fusion,the interspinous fixation device(BacFuse)model M2 after endoscopic decompression,and the interspinous fixation(BacFuse)model M3 after endoscopic-assisted interbody fusion were established.The same stress was applied to the upper surface of the L4 vertebral body in the four groups,and the lower surface of the L5 vertebral body was fixed and supported.The range of motion and the extreme Von Mises stress of the endplate bone and the posterior ligament complex of the vertebral body were analyzed under six working conditions of flexion,extension,left/right bending,and left/right rotation. RESULTS AND CONCLUSION:(1)Compared with model M0,the range of motion value of model M1 increased significantly under six working conditions.Model M2 and model M3 had a significant reduction in range of motion.(2)Compared with model M0,the maximum stress of the vertebral body in model M1 did not change significantly under the six working conditions.The maximum stress at the rear of the M2 vertebral body increased significantly.(3)Compared with model M1,the maximum stress of model M3 did not change significantly under the six working conditions.Compared with model M2,the maximum stress of model M3 decreased significantly.(4)Compared with the model M0,the extreme Von Mises stress of the L4 and L5 endplates of the model M1 was significantly increased.The extreme Von Mises stress in L4 and L5 endplates of models M2 and M3 decreased slightly.Compared with model M1,the Von Mises stress of the bone under the L4 and L5 endplate of models M2 and M3 was significantly reduced.(5)It is concluded that the implantation of BacFuse can effectively reduce the bone stress under the endplate during simple interbody fusion,decrease the risk of cage subsidence,diminish the risk of facet joint fracture on the decompression side,and provide a good stable environment for interbody fusion.The placement of an intervertebral fusion cage can reduce the stress of the root of the spinous process,which is beneficial to decrease the risk of fracture of the root of the spinous process.