1.Transgenic Production of Polyunsaturated Fatty Acids in Mammalian Cells
Ping KONG ; Zhuo DU ; Bo TANG ; Qingyong MENG ; Ning LI
Progress in Biochemistry and Biophysics 2006;0(11):-
Linoleic acid (C18∶2n-6) and ?-linolenic acid (C18∶3n-3) are found widely in fungi, plants and some lower animals. However, they can not be synthesized in mammals due to lack of △12 and ?-3 fatty acid desaturases. To enable endogenous production of essential fatty acids in mammalian cells, here the stable expression of a Caenorhabditis elegans gene FAT-2 encoding △12 fatty acid desaturase in CHO cells was reported. First, the FAT-2 coding sequence was cloned by RT-PCR. To facilitate high level synthesis of heterogeneous protein, the codon usage of the fatty acid desaturase genes was optimized according to the codon preference of mouse by site-directed mutagenesis, 2 synonymous mutations were introduced into FAT-2 gene by overlapping PCR. The codon-modified gene was finally fused to pBudCE4.1 vector (Invitrogen) under the control of CMV promoter. The expression vector pBudCE-FAT2 was linearized with NheⅠ, and then transfected CHO cells, the cells were under Zeocin selection for nine days and then propagated, then the transfected cells were harvested. The genome and total RNA were isolated for PCR and Norhern blot ananlysis. The results revealed that FAT-2 gene has been integrated into the genome of CHO cells and expressed properly. Fatty acids of total cellular lipids were analyzed by gas chromatography. The results indicate that the expression and function of △-12 fatty acid desaturase resulted in accumulation of linoleic acid. The levels of linoleic acid in transgenic cells were 2.4-fold higher than those in wild-type cells. The moderate linoleic acid in CHO cells was derived from cell culture media uptaken by cell membrane. The results demonstrate that a heterogenous desaturase gene can function well in mammalian cells and prove that transgenic approach is an efficient strategy for changing fatty acid composition of mammals.
2.Bcr-abl gene positive essential thrombocythemia treated with imatinib: one case and literatures review
Huanxun LIU ; Pengcheng WANG ; Yan LING ; Xin DU ; Jiacai ZHUO
Journal of Leukemia & Lymphoma 2008;17(2):131-132,135
Objective To observe the efficacy of imatinib on the treatment of bcr-abl positive essential thrombocythemia (ET). Methods A case of bcr-abl positive ET resistant to hydroxyurea (HU) treating with imatinib (200~400 mg/d) was reported and related literatures were reviewed. Results A case of bcr-abl positive ET was initially treated with 1.5~2.0 g/d HU, the platelet count decreased to 562x109/L after 4 weeks; however, the platelet count increased to (1020~1330)×109/L treating with same dose of HU 16 months later. With the elevation of HU to 3.0 g/d, platelet count was still high(1290~1780)x109/L companied with the very low white blood cell count(0.3~0.9)×109/L. While treating with imatinib (400 mg/d) for 1 month,the platelet count decreased to 390×109/L and white blood cell count was 0.5×109/L; Furthermore, treating with 200×300 mg/d of imatinib, the platelet and white blood cell count recovered in normal after 1 month,and bcr-abl fusion gene negative 2 months later. Conclusion Imatinib may be the effective targeting drug for the bcr-abl positive ET, and the bcr-abl positive ET is sensitive to low dose imatinib.
3.LISS locking plate and common plate in the repair of intra-articular proximal tibial fractures:choice of optimal implant
Yuxi DU ; Nianxi LIU ; Zhihui NIU ; Zhuo LIU
Chinese Journal of Tissue Engineering Research 2015;(35):5669-5673
BACKGROUND:The proximal tibial fractures within joint are a common type of fracture, and clinical treatment is difficult. The implant internal fixation is a commonly used method of treatment. However, the choice of the optimal implant internal fixation is always the key problem in clinical research. OBJECTIVE:To explore the curative effect of different implant internal fixation for the treatment of intra-articular proximal tibial fractures and to obtain the better treatment through the comparative analysis. METHODS:A total of 32 patients of intra-articular proximal tibial fracture, who were treated with different implant internal fixation for the treatment of proximal tibia, were selected from the Department of Orthopedics, People’s Hospital of Zezhou County of Jincheng City and Second Affiliated Hospital of Shanxi Medical University from September 2010 to September 2013. These patients were divided into two groups according to fixation methods (n=16). Observation and control groups received LISS locking plate and common plate, respectively. The amount of bleeding, operation time and incision length were observed in both groups. Patients were fol owed up for 1 to 12 months after treatment so as to record fracture healing time, ful weight bearing time and excel ent and good rate and to observe the incidence of complications, including joint stiffness, loosening, nonunion and infection. RESULTS AND CONCLUSION:Incision length and operation time in the observation group were significantly shorter than that in the control group, and the amount of bleeding was significantly lower in the observation group than that of control group (al P<0.05). The healing of fracture and the ful weight bearing time were significantly shorter in the observation group than that in the control group (both P<0.05). The incidence of complications was significantly lower in the observation group (13%) than in the control group (46%;P<0.05). The total excel ent and good rate of repair was significantly higher in the observation group than in the control group (94%, 69%, P<0.05). These results confirm that compared with the traditional plate fixation, internal fixation with LISS locking plate for treating intra-articular proximal tibial fractures can get better therapeutic effects, greatly reduces the injuries to soft tissue and fracture blood supply, promotes fracture healing and restores joint stability.
4.Imatinib combined with hematopoietic stem cell transplantation or chemotherapy for bcr-abl positive acute lymphoblastic leukemia
Huanxun LIU ; Jiacai ZHUO ; Xin DU ; Qingxiang MENG ; Ming LI
Journal of Leukemia & Lymphoma 2008;17(3):202-204
Objective To investigate the efficacy of imatinib combining with allogeneic hematopioetic stem cell transplantation or chemotherapy for bcr-abl positive acute lymphoblastic leukemia (ALL). Methods 12 cases were diagnosed on morphology, cytochemistry, immunophenotype and bcr-abl fusion gene. The induction is imatinib (400 mg/d) combining chemotherapy. 8 cases accepted allogeneic hematopoietic stem cell transplantation after complete remission (CR). If bcr-abl became positive, the patient was treated with imatinib (400~600 mg/d). 3 cases were tested with imatinib alternating chemotherapy after cr. Results 11 patients gained CR, CR rate 91.7 %; 5 patients (41.7 %) became bcr-abl negative through 2 courses induction. 3 cases relapsed after transplantation. 2 cases relapsed in imatinib combining chemotherapy group. The median remission interval is 16 months (imatinib combining transplantation group) and 10 months (imatinib combining chemotherapy group) (P <0.01) respectively. The median survival time is 18 months (imatinib combining transplantation group), and the other group (imatinib combining chemotherapy) is 12 months (P <0.01). Conclusion Imatinib combining chemotherapy achieved high CR rate for the bcr-abl positive ALL. Imatinib combining allogeneic hematopoietic stem cell transplantation is superior to imatinib combining chemotherapy for CR patients.
5.Reflections on cancer treatment
Qizhong ZHUO ; Zemin WANG ; Yanlin DU ; Yanjun Lü
International Journal of Traditional Chinese Medicine 2012;34(9):810-811
Incidence of cancer is increasing year by year,diagnostic techniques are continually advanced,and the numbers of anti-cancer drugs are growing,but the efficacy has not been fundamentally improved.Surgery,chemotherapy and radiotherapy can only kill the part of mature tumor cells,but can not kill the cancer stem cells,and eventually leading to tumor recurrence.While acupuncture,qigong,psychology comprehensive treatment can inhibit tumor growth and reduce metastasis and recurrence,improve quality of life and prolong survival.We should update our recognition,to carry out the study on the treatment of tumor by TCM with large sample and multiple centers,to demonstrate its low toxicity,low costs and efficiency.
6.Improvement and evaluation of chronic bronchitis modeling methods in mice
Xiuting DU ; Liang LUO ; Wanjun XIE ; Zhixun XIAO ; Guifeng ZHUO ; Ning SU
Chinese Journal of Pathophysiology 2015;(9):1724-1728
AIM:To explore a more accurate and reliable pathological model of the chronic bronchitis , which has improved from the former single-factor modeling method of the disease .METHODS:The mice in complex group were treated with lipopolysaccharide ( LPS) by tracheal injection on the 1st day and nasal drops on the 14th day, and from the 2nd day to 30th day, the animals were given passive smoking and sulfur dioxide ( SO2 ) inhalation ( except on the 14th day).The mice in SO2 group were exposed to SO2 2 min per day, while in smoking group, the mice were exposed to smoke for about 1 h per day (4 cigarettes each time until one pack of cigarettes were burning up ).In LPS group, the mice had tracheal injection of LPS on the 1st day and nasal drops of LPS on the 14th day and 30th day.Every modeling process las-ted for 30 days.After modeling, the improvement of chronic bronchitis model was evaluated by testing the general condi-tions of the mice , analyzing leukocyte count in bronchoalveolar lavage fluid ( BALF ) , and observing the morphological changes of the bronchial and lung tissues .RESULTS:After modeling, the mice in every model group experienced symp-toms including wet nose, cough, dry and lusterless hair, arched back and curled-up body, showing inactive, and slow down in response .The mice in complex group gained the lowest weight compared to other groups .From each model group , the inflammatory cells infiltrated evidently around the bronchial walls , especially in the bronchial cavity , and the mucilage secretion in the airway increased .The total number of leukocytes in BALF increased significantly in complex group .The in-flammatory cell count in the lung tissue indicated that the mice in complex group had significantly higher levels of inflamma -tory cell infiltration.Besides, the comparison between smoke group and LPS group was statistically significant .CONCLU-SION:Smoking, SO2 inhalation and LPS injection induce bronchial lung disease in mice , and the complex chronic bron-chitis mouse model is a better model with the pathological changes of bronchus , lung tissue and BALF , and pathogenesis of chronic bronchitis .
7.Changes of content of monoamine neurotransmitters and expression of neurotrophic factors in brain regions of rat models of anxious depression
Hongqing ZHAO ; Yuanshan HAN ; Zhuo LIU ; Qing DU ; Qin YANG ; Pan MENG ; Hui YANG ; Yuhong WANG
Acta Laboratorium Animalis Scientia Sinica 2017;25(4):373-379
Objective To study the content of monoamine neurotransmitters and neurotrophic factor in the hippocampus, amygdala and prefrontal cortex in anxious depression rats, and explore the possible pathogenesis.Methods 60 SD rats were randomly divided into normal group, vehicle group, anxiety group, depression group, and anxious depression group, 12 rats in each group.Chronic restraint stress combined with corticosterone injection was used to establish anxiety and depression model, the modeling time was 21 d.After modeling, elevated plus maze test, open field test, and forced swimming test were used to evaluate the anxiety and depression-like behavior, HPLC-ECD was used to detect the content of 5-HT, NE, and DA in the hippocampus, amygdala, and prefrontal cortex of rats.Western-blotting was used to detect the expression of BDNF and NT-3 in rats.Results Rats in anxious depression model group were comparable to the anxiety group in time and frequency entering open arm time, and number of locomotor activity in open field, and it had a significant difference when compared with the control and depression groups (P<0.01 or P<0.05).Immobile time in anxious depression model rats was increased significantly when compared with the control and anxiety groups (P<0.01).Meanwhile, compared with the control group, 5-HT in hippocampus and 5-HT, NE in amygdala or prefrontal cortex were significantly decreased in the depressive rats with anxiety (P<0.01 or P<0.05).Moreover, the content of BDNF and NT-3 was significantly decreased in each brain regions compared with the control group (P<0.01 or P<0.05), and BDNF levels were obviously decreased compared with the anxiety group (P<0.05).Conclusions Rats of anxious depression have significant anxiety and depression-like behaviors.Its mechanism may be associated with the down-regulation of monoamine neurotransmitters and neurotrophic factors BDNF and NT-3 in hippocampus, amygdala, and prefrontal cortex region.
8.Effects of Baishile Capsules on Hippocampal PI3K Signaling Pathway in Chronic Mild Unpredictable Stress Rats
Pan MENG ; Qing DU ; Xiuli ZHANG ; Zhuo LIU ; Yun XIANG ; Lin LIU ; Yuhong WANG
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(8):41-45
Objective To observe the effects of Baishile Capsules on the hippocampal PI3K signaling pathway in chronic mild unpredictable stress (CUMS) rats; To discuss its mechanism of action for anti-depression. Methods SD rats were randomly divided into 6 groups, control group, model group, fluoxetine group, Baishile Capsules high-, medium- and low-dose groups, with 10 rats in each group. The cums depression model was established, and gavage for medication was conducted at the same time for 21 days in a row. The behavioral changes of rats in each group were detected by the novel feeding experiment and Open-field experiment; monoamine neurotransmitter in serum were detected by ELISA; the expressions of PI3K, AKT and SGK1 in rat hippocampus were detected by immunofluorescence and Western blot test. Results Compared with the control group, feeding latency of rats in model group was significantly prolonged (P<0.01), horizontal and vertical activities were significantly reduced (P<0.01); the content of serum monoamine neurotransmitter decreased (P<0.01); the expressions of PI3K and AKT in hippocampus decreased, while the expression of SGK1 increased (P<0.01, P<0.05). Compared with the model group, Baishile Capsule high- and medium-dose groups can significantly shorten the incubation period of feeding rats (P<0.05), and increase the number of horizontal and vertical activities (P<0.01); the 5-HT and NE levels in serum were significantly elevated (P<0.05, P<0.01); the expressions of PI3K, AKT in hippocampal increased; the expression of SGK1 decreased (P<0.05, P<0.01). Conclusion Baishile Capsules can alleviate the depression like behavior in rat models, and regulate key factors of hippocampal PI3K signaling pathway, so as to exert antidepressant effects.
9.Integrating problem-based learning with mobile learning on virtual hospital platform
Hong ZHANG ; Xufang SUN ; Weikun HU ; Hao DU ; Wei WANG ; Zhuo CHEN
Chinese Journal of Medical Education Research 2013;(10):1057-1059,1060
Mobile learning has the characteristics of mobility,timeliness,network and virtu-ality. We developed virtual hospital suitable for mobile devices and organized students in to a network. Through early training,clinical case screen,learning objective fix,learning process and results evaluation,mobile devices were employed to do problem-based learning,which is conducive to the integration of theory with practice,the shifting from discipline-centered to system-based courses and the improving of utilization rate of learning resource.
10.Practical application of improved lateral radiography of the hip joint
Lei XUE ; Xu SHANG ; Zhuo LIU ; Xiangke DU ; Hao LIANG ; Weiwei QI
Journal of Practical Radiology 2017;33(9):1443-1446
Objective To design a simple and easy to operate new position radiography as same effect as the ordinary lateral position of the hip joint.Methods Using different angles for bone model radiography, the tilt angle of X-ray and the bone model were measured in order to obtain that the femoral head and neck was fully displayed.30 subjects with hip joint disease underwent routine and improved lateral radiography of the hip joint were enrolled, and the actual operations and image quality of the two methods were compared.Results The tilt angle of X-ray was 35°-45°,and the tilt angle of the bone model was 60°-70°, and the score was 3.The new lateral radiography of the hip joint was feasible,the display rate of the articular surface and joint space were 96.7%,the display rate of the femoral head and neck all were 100%,the display rate of the greater trochanter of femur was 80%,the display rate of the lesser trochanter was 100%.Conclusion The improved method of hip joint lateral position of X-ray can better show the femoral head, neck and the rest of the bone.